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Annals of Tropical Medicine 
and Parasitology 




THE UNIVERSITY OF LIVERPOOL 


Annals 

OF 

Tropical Medicine and 
Parasitology 


ISSUED BY THE 

Liverpool School of Tropical Medicine 


volume II 

(March a, 1908 to May 12, 1909) 


IVith twenty-three Plain and Coloured Plates 


LIVERPOOL: AT THE UNIVERSITY PRESS 
LONDON : ARCHIBALD CONSTABLE & CO., 16 JAMES STREET. HAYMARKET 




CONTENTS 


No. i 

Boyce, Sir Ruber r; and Breinl, Anton. 

Atoxyl and Trypanosomiasis ........ 

Manson, Sir Patrick. 

My Experience of Trypanosomiasis in Europeans and its Treatment by 
Atoxyl and other drugs ........ 


No. 2 

Kinghorx, Allan ; and Montgomery, R. Eustace. 

Reports of the ‘ Sleeping Sickness ’ Expedition to the Zambesi, for the 
year 1907-1908 ......... 

Montgomery, R. Eustace; and Kinghorn, Allan. 

A Report on Trypanosomiasis of Domestic Stock in North-Western 
Rhodesia 

Hadjimichalis, M.; and Cardamatis, Jean P. 

Report of the work of the Greek Antimalaria League during the year 

l 9°7 .. 


No. 3 

Rogers, Leonard. 

A Peculiar Intralobular Cirrhosis of the Liver produced by the 
Protozoal Parasite of Kala-azar .... 

Looss, Dr. A. 

What is ‘ Sckistosomum mansoni ’ Sambon 1907 r 
Ross, E. H. 

The Prevention of Dengue Fever ..... 

Salvin-Moore, J. E.; Breinl, Anton ; and Hindle, Edward. 

The Life History of trypanosoma lezvisi . 


PAGE 

1 

33 

S3 

97 

T 33 

H7 

153 

i93 


*97 



CONTENTS 


Moore, Benjamin ; Nierenstein, Maximilian ; and Todd, John Lancelot, page 

Notes on the Effects of Therapeutic Agents on Trypanosomes in respect 
to (#) Acquired Resistance of the Parasites to the Drug, and 
( b ) Changes in Virulence of the Strains after Escape from the Drug 221 

Nierenstein, M. 

Observations on the Acidity and Alkalinity of the Blood in Trypanosome 


Infections .......... 227 

Breinl, Anton; and Hindle, Edward. 

Contributions to the Morphology and Life History of PiropLisma ranis 233 

Nierenstein, M. 

Comparative Chemo-therapeutical Study of Atoxvl and Trvpanocides. 

Part I .......... 249 

d’Emmekez de Charmoy, M. 

On Three New Species of Culex collected during the Anti-malarial 

Campaign in Mauritius in 1908 ...... 257 


No. 4 

Moore, Benjamin; Nierenstein, M.; and Todd, John L. 

Concerning the Treatment of Experimental Trypanosomiasis. Part II 265 


Orpen, Dr. R. W. 

An Unusual Case of Goundou ....... 289 

Simms, Henry. 

Sub-Drainage as Applied to the Anti-malarial Campaign on the 

Isthmus of Panama ........ 291 

Theobald, F. V. 

A New Culicid Genus ......... 297 

Jeans, Frank. 

The Inflicted Talipes of the Chinese ...... 299 

Broden, A.; et Rodhain, J. 

Contribution a Petude de Porocephalus moniliformis . . . . 303 

Stephens, J. W. W. 

A New Human Nematode, Strongylus gibsoni, n. sp. . . • 3 J 5 



CONTENTS 

Stephens, J. W. W. page 

On the Supposed Occurrence of Filaria ivimitis in Man . . -317 

Breinl, Anton; and Hindle, Edward. 

A New Porocephalus (Porocephalus cercopitheci , n. sp.) . . *321 

Nierenstein, M. 

Comparative Chemo-therapeutical Study of Atoxyl and Trypanocides. 

Part II . 323 

Nierenstein, M. 

Chemical Notes on Atoxyl . . 329 

Sergent, Le Dr. Edmond. 

Notesur le role desTabanides dans la Propagation des Trypanosomiases 331 


No. 5 

Montgomery, R. Eustace; and Kinghorn, Allan. 

On the Nomenclature of the Mammalian Trypanosomes observed in 

North Western Rhodesia . . -333 

Breinl, Anton. 

Experiments on the Combined Atoxyl-Mercurv Treatment in Monkeys 

Infected with Trypanosoma gamble use . . . -345 

Marsden, Prosper H. 

Drugs from the Congo . . . . . -353 

Ross, E. H. 

A Gregarine Parasitic in the Dog-Flea, Ctenocephalus serraticeps . -359 

Breinl, Anton ; and Nierenstein, M. 

The Action of Aryl-Stibinic Acids in Experimental Trypanosomiasis 365 

Breinl, Anton; and Annett, H. E. 

Short Note on the Mechanism of Haemolysis in Piroplasmosis cams . 383 

Montgomery, R. Eustace; and Kinghorn, Allan. 

Gland Puncture in the Diagnosis of Animal Trypanosomiasis . . 387 

Stephens, J. W. W. 

Observations on the Hooklets of Cysiicercus cellvlosae in Man . . 391 




INDEX 




INDEX 


Index of Authors . iii 

General Index . iii 


Index of Genera and Species new to Science ... ix 


INDEX OF AUTHORS 


PAGE 

Boyce, R. i 

Breinl, A. i, 233, 321, 345, 365 

Broden, A. 303 

Cardamatis, J. P. 133 

d’Emmerez de Charmoy, M. 257 

Hadjimichalis, M. 133 

Hindle, E. 233, 321 

Jeans, F. 299 

Kinghorn, A.53, 97, 333 

Looss, A. 153 

Manson, P. 33 

Marsden, P. H. 353 

Montgomery, R. E.53, 97, 333 


PAGE 

Moore, B.221, 265 

Nierenstein, M., 221, 227, 249, 265, 323, 

329.36s 

Orpen, R. W. 289 

Rodhain, J. 303 

Rogers, L. 147 

Ross, E. H.193, 359 

Sergent, Ed. 331 

Simms, H. 291 

Stephens, J. W. W.315, 317, 391 

Theobald, F. V. 297 

Todd, J. L. 221, 265 


GENERAL INDEX 


PAGE 


Afridol blue in trypanosomiasis. 11 

„ violet in trypanosomiasis . 11 

Amino phenyl stibinic acids . 368 


et seq. 


„ „ dosage of 378 

„ ,, prepara¬ 

tion of 379 

Aniline dyes in treatment of experi¬ 
mental trypanosomiasis . 271 

Anophelinae of Mauritius. 260 

Anophelines in Greece . 139 

Antimony in treatment of trypanoso¬ 
miasis. 31 

„ „ (human) ... 44 

Arsenic and brilliant green in trypano¬ 
somiasis . 13 

,, dyes in treatment of try¬ 
panosomiasis . 26 

,, in the treatment of fly disease I 
,, use of by Bruce for fly disease 2 

n9 „ Lingard for Surra ... 3 

„ Livingstone for fly 

disease . 1 


PAGE 


Arsenious acid and trypan-red in try¬ 
panosomiasis . 13 

Aryl-stibinic acids in trypanosomiasis 365 
Atoxyl (acetylated) in treatment of 

experimental trypanosomiasis . 273 

Atoxyl, Amido group of. 325 

„ and mercury in trypanosomiasis 31 

,, „ trypanosomiasis. 1 

„ Chemo-therapeutical Study of 323 

„ combination with proteins ... 323 

„ Composition of. 28 

„ derivatives, Mode of action 253 

,, in skin diseases . 14 

„ „ sleeping sickness. 25, 26, 27 

„ „ trypanosomiasis . 18, 28 

(human) 

_ 33 , 42, 4 8 

Atoxyl-Mercury in trypanosomiasis 31 
„ ,, treatment in monkeys 345 

„ „ „ Cures effected by 351 

„ in segregation 

camps... 347 

„ „ „ mode of action 351 














































IV 


PAGE 

Atoxyl, mode of action . 250 

„ Parafuchsin in . 330 

„ Permeability of meninges to 327 
„ powder, Inorganic arsenic in 330 

„ -resistance . 29, 31 

„ secreted in urine . 326 

,, specific action of its amido 

group . 255 

,, toxicity of . 14 

treatment of trypanosomiasis 

experimentally . 272 

„ Use of, in trypanosomiasis 

16, 17, 27 

,, Water of crystallisation of. 329 

„ (yellow), Poisonous properties 

of . 33° 

Atylotus spp. transmit Debab . 331 

Autolysin not present in infection with 

Piroplasma cants . 385 

Baudoinville, Sleeping sickness at ... 85 , 

Benzidine colours in trypanosomiasis, 

i°, 11 ! 

Biringer, F. Use of atoxyl in skin j 

diseases . 14 j 

Blumenthal, F. On the toxicity of 

atoxyl . 14 

Boyce, R., and Breinl, A. On atoxyl 

and trypanosomiasis . 1 

Braid. On the use of Arsenic for fly 

disease . 2 

Breinl and Todd. On use of atoxyl in 

trypanosomiasis . 23 

Brilliant green in trypanosomiasis. 9 

Broden and Rodhain. On atoxyl in 

sleeping sickness . 26 

Browning, C. On atoxyl and para¬ 
fuchsin in trypanosomiasis. 11 

Bruce. On arsenic for fly disease. 2 ; 

Chichester, and Manson, P. On use j 
of arsenic in cattle trypanosomiasis 7 

Chinese, The inflicted talipes of. 299 

Cinchona alkaloids, Use of* by Lingard, 

for Surra . 3 

Cirrhosis, Atrophic, in Bengal. 152 

„ of liver in kala-azar . 147 1 

„ „ malaria. 151 

Colouring matters in trypanosomiasis 10 

Congo, Drugs from. 353 

„ 5 . mansoni in . 156 

Cook. On use of atoxyl in sleeping j 
sickness . 27 j 


PAGE 


Culex arboricollis , n. sp. 257 

„ * syn.'of Nausttadina 

arboricollis . 297 

Culcx fatigans conveys Dengue fever 193 

,, fozvleri , n. sp. 258 

„ „ syn. of Grabhamia 

fowleri . 297 

„ ronaldi , n. sp. 259 

„ „ syn. of Culcx micro - 

annulatus (Theo.) ... 297 
„ tigripes , carnivorous larvae of 262 

Culicinae of Mauritius . 261 

Cysticercus cellulosae (human) hooklets 

.of . 391 

„ ,, (pig) hooklets of 394 

Debab transmitted by Atylotus , spp.... 331 
Dengue fever conveyed by Culex 

fatigans . 193 

„ ,, exterminated by anti¬ 
mosquito campaign... 194 

„ ,, prevention of . 193 

Dogs, trypanosomiasis of.118, 124 

Drugs from Congo. 353 

Egypt, S. mansoni in . 156 

Ehrlich and Shiga. On use of trypan- 

red in trypanosomiasis. 8 

„ On composition of atoxyl ... 28 

„ On use of benzidine colours 

in trypanosomiasis . 10 

Filaria immitis , supposed occurrence 

in man . 317 

„ papillosa in Rhodesia . 107 

Flea of dog, gregarine in . 359 

Fly disease, treatment by arsenic. 1, 2 

„ treatment by quinine ... 2 

Fourneau. On composition of atoxyl 28 

Franke, E. On trypan-red in try¬ 
panosomiasis. 9 

,, On trypan-red-arsenic in 

trypanosomiasis . 13 

Fritsch. On structure of female 

5 . haematobium . 159 

Gland palpation, Statistics of, in 

Luapula (Rhodesia) 82 

„ „ Statistics of in 

Nyassaland 83 

Gland puncture in trypanosomiasis 

(animal) . 387 

Glandular enlargement in Rhodesia 66 

Glossina fusca on Lake Nyassa . 69 

,, „ on Lake Tanganyika ... 66 






















































V 


PAGE 


Glossina palpalis , distribution in 

Rhodesia.60, 64 

,, „ on Lake Mweru. 85 

„ „ „ Tanganyika, 66,85 

„ spp., possible carriers of 

sleeping sickness . 65 

Goundou, Curvature of tibia in . 290 

„ Unilateral (illustrated) ... 290 

„ Unusual case of . 289 

Grabhamia fowleri = Culex fowleri ... 297 

Greece, Anophelines in . 139 

„ Deaths from malaria in . 133 

„ Quinine administration in ... 140 

Greek antimalaria league report. 133 

Gregarina ctenocephali cants . 363 

Gregarine in dog-flea . 359 

Haemolysis in infection with Piro- 

plasma cants . 383 

Halberstadter. On trypan-red in try¬ 
panosomiasis . 9 

Hollebecke. On atoxyl in sleeping 

sickness . 27 

Isolysin not present in infection with 

Piroplasma canis . 385 

JakimoflF. On atoxyl in trypano¬ 
somiasis . 27 f 

Kabinda, Sleeping sickness in . 77 

Kala-azar, Cirrhosis of the liver in ... 147 
Katanga, Congo Free State, Tsetse 

flies in . 81 

Kisale Lake, Sleeping sickness around 77 
Koch. On Afridol blue in trypanoso¬ 
miasis . 11 

„ ,, violet in trypano¬ 
somiasis . 11 

„ atoxyl in sleeping sickness 25 

„ parafuchsin in trypanoso¬ 
miasis . 11 

,, pararosaniline in trypano- 


PAGE 


Lingard. On arsenic for Surra. . 3 

Livingstone, D. On arsenic for tsetse 

fly disease . 1 

Luapula (Rhodesia), Sleeping sickness 

in . 84 

„ „ Statistics of gland 

palpation in... 82 

Lyperosia and cattle trypanosomiasis... 129 

„ sp. and trypanosomiasis. 59 

Maghalhaes. On arsenic and brilliant 

green in trypanosomiasis. 13 

Malaria (anti) league in Greece . 133 

„ at Marathon. 138 

„ Deaths from, in Greece . 133 

„ -transmitting mosquitoes in 

Mauritius . 262 

Malarial (anti) campaign in Panama 291 

„ cirrhosis of liver . 151 

Manson, P., and Chichester. On 
arsenic in cattle trypano¬ 
somiasis . 7 

,, On atoxyl in trypanoso¬ 
miasis (human) . 33 

Marathon, Malaria at . 138 

„ 4 Splenic index 9 at . 138 

Martin. On atoxyl in sleeping sick¬ 
ness . 27 

Mauritius, Malaria - transmitting 

mosquitoes in...... 262 

„ Mosquitoes of . 260 

Mendel, F. Use of atoxyl intrave¬ 
nously . 14 

Mercury and atoxyl in treatment of 

trypanosomiasis . 31 

„ perchloride and atoxyl in 

trypanosomiasis 277 
„ „ in treatment of 

Ngana . 7 

,, „ in treatment of 


somiasis . 11 

Kopke. On atoxyl in sleeping sickness 27 

‘ Latent * bodies of 7 . equiperdum . 202 

„ „ 7 . gambiense . 202 

Laveran and Mesnil. On Sodium 
arseniate in 
Ngana . 7 

„ „ trypan-red in 

trypanosomia¬ 
sis. 9 

,, On trypan-red and arsenious 

acid in trypanosomiasis. 13 


i trypanosomiasis 

(human) . 43 

,, „ Use of, by Lin¬ 
gard, for Surra 3 

Mesnil, Nicolle and Aubert. On 
arsenic and dyes in trypano¬ 
somiasis . 26 

Mesnil, Nicolle and Aubert. On 
| benzidine colours in trypanosomiasis 10 

I Moliro, Sleeping sickness at . 85 

Moore, E. J. Treatment of tsetse- 
fly disease by sodium arseniate. 7 





















































VI 


PAGE 

PAGE 

Moore, Nierenstein and Todd. On 
composition of atoxyl . 

28 

P. moniliformis. Experimental infec¬ 
tion of mammals and 


Mosquito (anti) campaign, Extinction 


man with adults. 

00 

0 

to 

of Dengue fever by . 

' 9 + 

„ Experimental infec- 


Mosquitoes of Mauritius . 

260 

tion of snakes with 


Mouindu (drug) . 

353 

larvae . 

3 ii 

Muindu (drug) . 

353 

„ larval forms in 

Mweru lake, Glossina palpalis on. 

85 

monkeys . 

303 

Newsteadina arboricollis = C. arbori- 


,, Life history of. 

303 

collis . 

297 

Potassium antimony tartrate, Try- 

Ngana not transmitted by Taba- 


panocidal action of. 

365 

nidae . 

33 i 

Prophylaxis of sleeping sickness... 88 et 

seq. 

„ treatment by human serum ... 

7 

Quinine administration in Greece. 

140 

„ „ mercury per- 

chloride . 

7 

„ used by Ranking for fly 

disease . 

2 

,, ,, silver salts . 

7 

Ranking. On quinine for fly disease... 

2 

„ ,, sodium arseniate 

7 

Rhodesia, Glandular enlargements in 

66 

„ „ sodium cacody- 


,, Glossina palpalis in .60, 64 

late . 

7 

„ T. dimorphon ? in . 

58 

Nissle. On trypan-red in trypanoso- 


„ T. vivax in. 

58 

miasis . 

q 

Trypanosomiasis in . 

58 

Nyassa, Lake, Gl. fusca on. 

69 

,, North-Eastern, Tsetse-flies 

Nyassaland, Statistics of gland palpa- 


of . 79, 80 

tion in . 

«3 

,, (N.W.), Trypanosomes of 

00 

to 

to 

„ Tsetse flies in . 

8l 

„ „ Trypanosomiasis of 


Panama, Anti-malarial campaign in... 

291 

dogs in . 

118 

Parafuchsin in atoxyl. 

330 

„ „ Trypanosomiasis of 


„ treatment of trypano¬ 


domestic stock in 

97 

somiasis (human) ... 

43 

„ „ F. papillosa in . 

107 

„ trypanosomiasis . 

11 

„ „ P. bovis in . 

107 

P aramphistomum conicum in Rhodesia 

107 

„ ,, P. conicum in . 

107 

Pararosaniline in trypanosomiasis. 

11 

,, „ T. dimorphon in 


Piroplasnta bovis in Rhodesia. 

107 

sheep in . 

Il 7 

,, rrt«zj,Autolysin not present 


„ ,, T. theilcri in. 

117 

in infection with ... 

385 

,, „ T. vivax in . 

112 

„ „ flagellated forms of 


Schild. Use of atoxyl in skin diseases 

H 

in blood . 

238 

S. haematobium eggs in tissues isolated 


„ „ Haemolysis in . 

383 

by potash . 

188 

„ „ Isolysin not present 


,, fate of miracidium ... 

1 77 

in infection with 

00 

<-n 

„ frequency of males in 


„ „ Morphology and life 


portal vein. 

161 

history of. 

233 

,, Fritsch on structure of 


Plimmer and Thomson :— 


female of . 

159 

On antimony in trypanosomiasis 

31 

„ habitat of mature 


On atoxyl and mercury com¬ 


worms . 

168 

pounds in trypanosomiasis. 

3 1 

„ lateral and terminal 


On benzidine colours in trypano¬ 
somiasis . 

11 

spined eggs in intes¬ 
tine . 

172 

Porocephalus cercopitheci , a new Poro- 
cephalus . 

321 

„ lateral and terminal 

spined eggs in same 


P. moniliformis adults in snakes. 

304 

female. 

161 























































Vll 


PACK 

S. haematobium lateral spinet! eggs in 

bile .. 170 

4>. lateral spined.eggs in 

liver . 168 

,, lateral spined eggs in 

lung . 170 

„ lateral spined eggs laid 

by young females ... 186 
„ lateral spined eggs of, 

always mixed with 
terminal spined eggs 
in rectum . 164 


75 

lateral spined eggs of 



not solely from 



rectum . 

164 

7 7 

mode of formation of 



egg with lateral spine 

160 

77 

mode of infection ... 

179 

77 

site of adults. 

180 

7 ? 

site of oviposition ... 

182 

77 

unfertilised females 



produce abnormal 



egg 8 . 

167 

S. mansotti. 

characters of eggs . 

15s 

77 

Geographical distribution 



of . 

156 

77 

Habitat of. 

156 

77 

in Congo Free State . 

156 

77 

in Egypt .. 

156 

77 

in West Indies . 

I 5 6 

77 

no special habitat . 

i 7 t 

99 

Sambon, 1907, What is ? 

153 1 

Serum, human, in treatment of Ngana 

7 1 

Silver salts 

in treatment of Ngana. 

7 ! 

Skin diseases, treatment by atoxyl. 

1+ 

Sleeping sickness around Kisale Lake 

77 

77 

at Baudoinville . 

8 5 | 

77 

Moliro. 

85 1 


Vua . 85 

endemic on Tangan¬ 
yika . 66 

Expedition to Zambesi 53 

in Kabinda. 77 

Luapula (Rhodesia) 84 
possibly carried by 

Glossina, spp. 65 

prophylaxis.88 et seq . 

treatment by atoxyl 

25,26,27 

treatment by sodium- 
antimonyl tartrate 366 


PACE 

Sleeping sickness treatment by sodium 


arseniate 8 

„ Vide trypanosomiasis 

(human) 

Sodium-antimonyl tartrate in sleeping 

sickness . 366 

„ arseniate, Use of by Laveran 
and Mesnii for 
Ngana . 7 


„ „ Use of, by Moore 

for tsetse-fly disease 7 
„ „ Use of by Thomas 

in treatment of 

Ngana . 7 

,, „ Use of by Thomas 

and Breinl in try¬ 
panosomiasis and 
sleeping sickness 8 
„ cacodylate in treatment of 

Ngana . 7 

‘ Splenic Index ’ at Marathon . 138 

Strongylus gibsoni , a new human nema¬ 
tode . 315 

Stomoxys and cattle trypanosomiasis... 129 

„ sp. and trypanosomiasis . 59 

Surra treatment by arsenic . 7 

„ „ cinchona alkaloids 7 

„ „ mercury per- 

chloride . 7 

Tabanidae transmit Debab not Ngana 331 
Talipes, The inflicted, of Chinese ... 299 

Tanganyika, Gl. jusca on . 66 

„ Glossina palpalis on ... 66, 85 

„ sleeping sickness, endemic 

on . 66 

Thiroux and d’Anfreville. On atoxyl 

in sleeping sickness. 27 

Thomas. On atoxyl iji trypanoso¬ 
miasis . 16, 17 

„ On sodium arseniate in 

treatment of Ngana . 7 

Thomas and Breinl:— 

On atoxyl in trypanosomiasis... 18 et seq. 
On sodium arseniate in trypanoso¬ 
miasis and sleeping sickness. 8 

On trypan-red in trypanosomiasis 9, 12 
Tragelaphus scriptus , Trypanosome 

of. 336 

Trypanocidal action of potassium- 

antimonyl tartrate. 365 

Trypanocidal amido group. 271 













































VIII 


PACK 


Trypanocidcs, Chemo- therapeutical 

study of . 323 

T. brucei , Acidity of blood serum in¬ 
creased in infections with . 227 

T. cazalboui . 340 

T. brucei , Changed virulence of when 

drug-proof . 224 

T. congolense . 339 

T. dimorphon group. 338, 339 

„ in cattle . 104 

,, in Rhodesia . 58 

„ in sheep in Rhodesia 

(N.W.) . 117 

„ Morphology of . 118 

T. rquiprrduni , Acidity of blood serum 
increased in infec¬ 
tions with . 227 

„ ‘ Latent bodies ’ of ... 202 

,, mode of multiplication 202 

T. evansi group .. 338, 339 

T. gambirnsr, alternating phases of 
presence and absence 

in blood . 200 

„ in monkeys, Treatment 

by atoxyl mercury ... 345 

„ ‘ Latent bodies ’of. 201 

„ mode of multiplication 200 

J. letvisi, amitotic division of nucleus 206 

„ Arbitrary distinction of 

sexual forms of . 212 

„ 4 Latent bodies ’ of . 210 

,, Life history of . 197 

„ Morphological details des¬ 
troyed by drying . 198 

„ Morphology of . 203 

T. nanum . 340 

T. pecaudi . 339 

T. sudanense . 339 

T. theileri in Rhodesia (N.W.). 117 

T. vivax . 58, 112, 340 

„ Morphology of. 122 

Trypanosome of Erythrea . 343 


„ Tragelapbus scriptus 336 

Trypanosomes, Changed virulence of 

when drug proof ... 224 

„ drug proof only for the 

animal in which 
they were made... 222 

,, Drug proof strains of 221 

„ Nomenclature of 

mammalian . 333 


PAGE 


Trypanosomes of North-Western 

Rhodesia . 338 

Trypanosomiasis, Acidity and alkalinity 

of blood in . 227 

„ and atoxyl. 1 

,, „ Lyperosia, sp.... 59 

„ Stomoxys , sp. ... 59 

„ (animal) gland punc¬ 
ture in . 387 

,, Aryl stibinic acids in 

treatment of . 365 

„ Combined treatment 

in . 31 

„ (experimental), 

aniline dyes used in 

treatment of . 269 

„ (experimental), 

inefficacious drugs 
used in treatment 
of . 269 


(experimental), 
treatment by atoxyl 272 
(experimental), 
treatment by atoxyl 


acetylated . 273 

(experimental), 

treatment of . 265 

(human), ascribed 

to bite on leg . 51 

(human), in females 50 
(human), treatment 

by antimony . 44 

(human), treatment 

by atoxyl. 33, 48 

(human), treatment 
by mercury per- 

chloride . 43 

(human), treatment 
by parafuchsin ... 43 

in Rhodesia . 58 

Mercury perchloride 
and atoxyl in treat¬ 
ment of. 277 


of cattle and Lypero¬ 
sia ... 129 
„ „ Stomoxys 129 

of cattle, mode of 

transmission . 128 

of cattle, treatment 
by arsenic by Chi¬ 
chester and Manson 7 
















































IX 


PACE 

Trypanosomiasis of dogs in Rhodesia 

(N.W.). 118.124 

„ of domestic stock in , 

N.W. Rhodesia ... 97 

„ treatment by anti- } 

mony . 31 ■ 

„ treatment by arsenic 

and brilliant green 13 

,, treatment by arsenic 

and dyes . 26 

,, treatment by arseni- 

ous acid and trypan- 

red. 15 

,, treatment by atoxyl 

16, 17, 18, 27, 28 

„ treatment by atoxyl j 

and mercury . 31 

„ treatment by benzi- , 

dine colours . 10 ' 

„ treatment by colour¬ 
ing matters . 10 1 

„ treatment by trypan- 

red. 8, 9, 11 1 

,, Trypan-red and 

mercury perchloride 
in treatment of ... 285 
Trypan-red and arsenious acid in try¬ 
panosomiasis. 13 

„ mercury perchloride 

in trypanosomiasis 285 


PACE 

Trypan-red in treatment of trypano¬ 
somiasis .8, 11 

Tsetse-fly and big game . 53, 67 

Tsetse-flies in Katanga, Congo Free 

State . 81 

„ Nyassaland. 81 

,, Rhodesia, North- 

Eastern . 79, 80 

Uhlenhuth, Gross, and Bickel. On 

atoxyl in trypanosomiasis . 28 

Van Campenhout. On atoxyl in 

sleeping sickness . 26 

Vua, Sleeping sickness at . 85 

Weber and Krause. On colours in 

trypanosomiasis . 10 

Wendelstadt and Felmer. On arsenic 
and brilliant green in trypanosomia¬ 
sis . 13 

Wendelstadt and Fellmer. On bril¬ 
liant green in trypanosomiasis. 9 

Wendelstadt. On trypan-red in try¬ 
panosomiasis. 9 

Wenyon. On benzidine colours in 

trypanosomiasis . 10 

West Indies, 5 . mansoni in. 156 

Wild game and tsetse-fly. 67 

Yakimoff. On trypan-red in trypano¬ 
somiasis . 11 

Zambesi, Sleeping Sickness Expedi¬ 
tion to. 53 


INDEX OF GENERA AND SPECIES NEW TO SCIENCE 


PAGE 


Culex arboricollis . 258 

,, fowleri . 258 

„ ronaldi . 259 

Gregarina ctenocephali canis . 363 


PAGE 


Newsteadina , nov. gen. 297 

Porocephalus cercopitheci . 321 

Strongylus gibsoni . 315 






































«. • 






Volume II 


March, 1908 


No. 1 


ANNALS 

OF 

TROPICAL MEDICINE AND 
PARASITOLOGY 


ISSUED BY 

THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE 


Editor 

Professor RONALD ROSS, Major I.M.S. (Ret.), D.P.H., F.R.C.S., 
D.Sc., LL.D., F.R.S., C.B. 


In Collaboration with 

J. W. w. STEPHENS, M.D., Cantab., D.P.H. 

R. NEWSTEAD, A.L.S., F.E.S. 

J. L. TODD, B.A., M.D., C.M. McGill. 

H. WOLFERSTAN THOMAS, M.D., C.M. McGill. 
ANTON BREINL, M.U.Dr. 

Professor Sir RUBERT BOYCE, M.B., F.R.S. 



C. Tinting & Co., Ltd. 

Printers to the University Press of Liverpool 
Victoria Street 



THE INCORPORATED 

LIVERPOOL SCHOOL OF TROPICAL MEDICINE 

(Affiliated with the University of Liverpool) 


Hon. President: Her Royal Highness Princess CHRISTIAN 
Hon. Vice-Presidents: The DUKE OF NORTHUMBERLAND, K.G. 
Earl Cromer, G.C.B. 


COMMITTEE 

Chairman : Sir ALFRED L. JONES, K.C.M.G. 

Vice-Chairman: Mr. WILLIAM ADAMSON, President Royal Southern Hospital 
Hon. Treasurer : Lord STANLEY, K.C.V.O. 


Vice-Chancellor Dale 

Sir W. B. Bowring, Bart. I 

Dr. CATON ) 

Professor Sir Rubert Boyce, ) 
M.B., F.R.S. L 

Professor Sherrington, F.R.S. j 
Dr. W. Alexander ) 

Dr. C. J. Macalister j 

Mr. J. O. Strafford 
Mr. T. F. Harrison ) 

Mr. Charles Livingston ) 

Mr. A. R. Marshall ) 

Mr. W. Roberts Ji 

Mr. Stanley Rogerson 
Mr. C. Booth (Jun.) 

Mr. A. F. Warr 
Mr. F. C. Danson 
Mr. George Brocklehurst 
M r. J. W. W. Danson 


University of Liverpool 

Council of University of Liverpool 

Senate of University of Liverpool 

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Staff 

Professor 

Major RONALD ROSS, C.B., F.R.S., F.R.C.S., D.Sc., 
D.P.H. (Ind. Med. Serv. retired). 

Lecturer 

J. W. W. STEPHENS, M.D. (Cantab.), D.P.H. 

Lecturer in Economic Entomology and Parasitology 
R. NEWSTEAD, A.L.S., F.E.S. 

Physicians 

Major RONALD ROSS,C.B.,F.R.S.,etc. ) 0 /c , 

C. J. MACALISTER, M.D., M.R.C.P. - " 

LLOYD ROBERTS, M.D., F.R.C.S. ) 

Surgeons 

VV. ALEXANDER, M.D., F.R.C.S. ) 

R. JONES, F.R.C.S. (EdiN.) \ Royal Southern 

G. P. NEWBOLT, M.B., F.R.C.S. J * 

Honorary Lecturers 
H. E. ANNETT, M.D., D.P.H. 

Professor of Comparative Pathology 
W. T. PROUT, M.B., C.M.G. 

C. E. WALKER, F.L.S. 

Assistant Director of Cancer Research 
Major C. L. WILLIAMS, M.D., D.P.H. 

(Ind. Med. Serv. retired) 

Runcorn Research Laboratories 
Director: ANTON BREINL, M.U.Dr.. Prag 
Research Demonstrator: M. NIERENSTEIN, Ph.D. 
Research Assistant : ED. HINDLE, A.R.C.S. (Lond). 

On Expeditions 

H. WOLFERSTAN THOMAS, M.D., C.M. 

ALLAN KINGHORN, M.B. 

R. E. MONTGOMERY, M.R.C.V.S. 

J. O. W. BARRATT, M.D., D.Sc. (Lond.) 
WARRINGTON YORKE, M.D. 

Dean 

Professor Sir KUBERT BOYCE, M.B., F.R.S. 
Laboratory 

JOHNSTON TROPICAL LABORATORY, LIVERPOOL 
UNIVERSITY 

Research Laboratory 
CROFTON LODGE, RUNCORN 

Hospital 

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Secretary's Office 

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NOTICE 


Regarding the Courses of Instruction proposed to be given by the 
Liverpool School of Tropical Medicine, and the Examinations for the 
Diploma of Tropical Medicine arranged to be held by the University 
of Liverpool during 1908 (subject to such alteration as may hereinafter 
be decided upon), 

Lent Term begins January 14. 

Lent Examination, March 23. 

Summer Term begins May 1. 

Summer Examination, July 13. 

Autumn Term begins October 1. 

Autumn Examination, December 14. 

The full Course of Instruction is open to all qualified medical men, 
and the examination to all students who have taken out this full 
course. 

Fee for the full Course of Instruction Ten guineas. 

Fee for the Examination Five guineas. 

Fee for the use of a School microscope during one term Fen 
shillings. 

For prospectus and further information, application should be made 
to the Dean of the Medical Faculty, University of Liverpool. 


The following have obtained the Diploma in Tropical Medicine of 
the University of Liverpool:- 

Diploma in Tropical Medicine 


Date of 
Diploma 

1906 Adie, Joseph Rosamond 

1907 Allan, Alexander Smith 
1907 Allwood, James Aldred 

1905 Anderson, Catherine Elmslie 

1906 Arnold, Frank Arthur 
1904 Augustine, Henry Joshua 
1906 Bate, John Brabant 

1904 Bennett, Arthur King 

1906 Bennetts, Harold Graves 

1907 Bond, Ashton 
1907 Branch, Stanley 

1905 Brown, Alexander 
1904 Bruce, William James 

1904 Byrne, John Scott 

1905 Caldwell, Thomas Cathcart 

1906 Carter, Robert Markham 
1906 Chisholm, James Alexander 

1904 Clayton, Thomas Morrison 

1906 Clements, Robert William 

1907 Collinson, Walter Julius 

1905 Critien, Attilio 

1904 Dalziel, John McEwen 
1907 Davey, John Bernard 
1904 Dee, Peter 
1907 Donaldson, Anson Scott 

1906 Dundas, James 

1906 Faichnie, Norman 

1907 Fell, Matthew Henry Gregson 
1907 Gann, Thomas William Francis 
1907 Graham, James Drummond 
1904 Greenidge, Oliver Campbell 

1904 Hehir, Patrick 

1907 Hiscock, Robert Carroll 

1905 Hooton, Alfred 

1905 Hudson, Charles Tilson 

1905 Illington, Edmund Moritz 

1906 Jeffreys, Herbert Cartel man 


Date of 
Diploma 

1907 Keane, Joseph Gerald 
1907 Kennan, Richard Henry 
1907 Kenrick, William Hamilton 
1904 Khan, Saiduzzafor 

1904 Laurie, Robert 

1907 Le Fanu, George Ernest Hugh 

1905 Macfarlane, Robert Maxwell 

1906 Mackenzie, Donald Francis 

1907 Mackey, Charles 

1904 Maclurkin, Alfred Robert 

1905 Maddock, Edward Cecil Gordon 
1907 Maddox, Ralph Henry 

1907 McCarthy, John McDonald 

1904 McConnell, Robert Ernest 

1905 Moore, James Jackson 

1904 Nicholson, James Edward 

1905 Nightingale, Samuel Shore 

1906 Pailthorpe, Mary F^lizabeth 
1906 Palmer, Harold Thornbury 

1906 Pearse, Albert 

1904 Philipson, Nicholas 

1905 Radcliffe, Percy Alexander Hurst 

1907 Raikes, Cuthbert Taunton 
1907 Ryan, Joseph Charles 

1906 Sampey, Alexander William 
1904 Sharman, Eric Harding 

190b Smithson, Arthur Ernest 
1906 Taylor, Joseph van Someron 
1906 Taylor, William Irwin 
1904 Thomson, Frank Wyville 

1906 Tynan, Edward Joseph 

1907 Vallance, Hugh 

1904 Walker, George Francis Clegg 
1906 Watson, Cecil Francis 
190O Willcocks, Roger Durant 
190(1 Williamson, George Alexander 
190:; Young, John Cameron 




EDITORIAL NOTICE 


By order of the Committee of the Incorporated Liverpool School 
of Tropical Medicine, the series of the Reports of the School, which 
have been issued since 1899, will be followed, from January I, 1907, 
by the Annals of Tropical Medicine and Parasitology, of which 
this is the first number of the second volume. 

The Annals are issued by the Committee of the School, and will 
contain all such matter as was formerly printed in the Reports—that 
is to say, accounts of the various expeditions of the School and of the 
scientific work done in its laboratories at the University of Liverpool 
and at Runcorn. Altogether twenty-one Memoirs, besides other 
works, have been published by the School since 1899, and of these 
ten, containing 519 quarto or octavo pages and 95 plates and figures, 
were published during the two years 1904 and 1905 ; and there is no 
reason to suppose that this rate of production by the workers of the 
School alone will diminish in the future. In addition, however, to 
School work, original articles from outside on any subject connected 
with Tropical Medicine or Hygiene may be published if found 
suitable (see notice on back of cover); so that, in all probability, not 
less than four numbers of the Annals will be issued annually. Each 
number will be brought out when material sufficient for it has been 
accumulated. 




ATOXYL AND TRYPANOSOMIASIS 


BY 

SIR RUBERT BOYCE, F.R.S., 

AND 

ANTON BREINL, M.U.D. Prag. 

(Received for publication January 20 tk, 1908 ) 

The brilliant discovery by the late Dr. Dutton in 1901 of 
the presence of trypanosomes in the blood of a patient under the 
care of Dr. Forde of Bathurst, to which he gave the name of 
Trypanosoma gambiense , the finding very shortly afterwards, by 
an expedition sent out to Africa under the auspices of the Royal 
Society and Colonial Office, composed of CASTELLANI, Bruce, 
NABARRO and LOW, that sleeping sickness was caused by the same 
parasite ( Trypanosoma gambiense ), stimulated investigation 
throughout the civilised world into the life history of this group of 
haematozoa, their mode of action in the blood and tissues of man and 
animals, and the effect of various drugs upon them. 

During the year 1907 very material progress has been made in 
the treatment of sleeping sickness, and it appears to us that the 
time is a suitable one in which to review the history of how arsenic 
and its compounds came to be employed, and to state the results of 
the treatment with this and allied drugs, in the light of the great 
experience gained in 1907. 

There is no doubt that for a very long time Arsenic has been 
looked upon as a remedy useful in trypanosomiasis in animals. Long 
before the nature of sleeping sickness was understood there existed 
much speculation with regard to the nature and treatment of Tsetse 
Fly disease in horses and cattle. First and foremost among those 
who suggested Arsenic as a means of treating this disease in animals 
stands the great observer and explorer Dr. Livingstone. 

David Livingstone, in a letter (dated March 22, 1858) to the 
British Medical Journal of May 1st, 1858, mentioned that it had 
occurred to him in about the year 1847-8 to use Arsenic in the disease 
which followed the bite of the tsetse fly. He mentions how he tried 
the drug on a mare. 

B 


Early observations 
upon the use of 
Arsenic in the treat¬ 
ment of Fly disease 



2 


Use of Arsenious 
acid and Sodium 
arseniate 


In thanking Mr. Braid for some remarks (published in the 
British Medical Journal) Livingstone states ‘that though my hopes 
are not sanguine, I still mean to try the remedy, if opportunity offers. 
Our instructions require us to examine the whole subject carefully, 
and the results will be communicated to the* Royal Society/ 

In the British Medical Journal of February 13, 1858, there 
appears a letter from Mr. Braid, dated from Manchester, February 6, 
1858, in which he says: ‘On reading the interesting facts 
communicated by Livingstone, one of the most notable is his narrative 
of the remarkable and fatal phenomena manifested in oxen and 
sheep from the bite of the tsetse fly. It immediately occurred to me 
that it would be highly interesting to institute some experiments 
with the view of discovering a remedy for this curious and fatal 
malady, and my mind immediately reverted to the prophylactic 
powers of arsenic against the poison of the most venomous reptiles 

Braid then quotes Dr. Honigbergers case of a fakir who was an 
Arsenic eater, and who ascribed to this reason his immunity to the 
bite of the serpent. 

Such at any rate was the source from which the idea developed of 
giving small doses of Arsenic to oxen bitten by the tsetse fly. 

Surgeon-Major RANKING had been struck with the similarity 
existing between the disease Surra in horses and mules and malaria, 
and recommended a treatment similar to that used in malaria, but 
although he freely used Quinine , we find no evidence that he 
employed Arsenic. 

We then come to the period of definite organised experiments, and 
foremost amongst investigators stands Bruce, who added an immense 
number of new facts to our very scanty knowledge of trypanosomiasis. 
BRUCE, in his classical report entitled ‘ Further Report on the Tsetse 
Fly disease or Nagana in Zululand ’ (May 1896), gives a very useful 
and detailed account of the effect of Arsenic as a curative and 
prophylactic agent. 

The experiments he set on foot demonstrated that Arsenic had a 
material effect, and that it was capable of driving haematozoa out of 
the blood; as Col. Bruce himself states, the drug ‘ undoubtedly 
markedly modified the course of the disease / As a prophylactic 
agent, he argued that if Arsenic modified the course of the disease, 



3 


it seemed probable therefore that the disease would be prevented 
altogether if the Arsenic were given for some time previous to the 
animals being exposed to the infection. 

From experiments set on foot to determine whether Arsenic had 
a prophylactic action, he concludes that it would be quite useless 
as a prophylactic agent, but that it was useful in prolonging life, 
and especially in the * Fly Country/ after the disease had begun. 

Following upon Bruce's observations come those of LlNGARD (1893). 

LlNGARD (Report on Horse Surra, vol. I, p. 104, 1893) mentions 
in connection with his experimental enquiry into the treatment of 
Surra, that Pottinger had tried Arsenic without any appreciable 
effect Lingard then conducted himself a series of experiments upon 
numerous animals with Arsenic compounds , Cinchona alkaloids , 

Arsenic alone, and also Mercury perchloride. The results showed in 
the case of Mercury perchloride that there was no diminution in the 
number or activity of the parasites in the blood, and that they 
continued to be present in the blood for three days following the 
injection of the drug. 

With regard to the treatment with Cinchona alkaloids and Combined treat- 
Arsenic y we quote Lingard’s own words ( 1 . c.):—* In the case of two thef 
animals, it was decided to attempt curative treatment, although it avoids 
was recognised that the animal had already been suffering from the 
disease for a period of at least twenty-three days. It was thought 
advisable to commence the treatment when the number of haematozoa 
in the blood should be on the decline, and accordingly on the 14th 
June, the haematozoa being few, treatment with Cinchona alkaloids 
and Arsenic in large doses was commenced. On the 20th the 
haematozoa disappeared from the blood, but the temperature and 
respirations remained above normal. The general symptoms of Surra 
increased up to the 24th; oedema and swelling of the limbs, sheath 
and under the abdomen increased in extent. The animal steadily 
losing condition, the muscles wasting, and the gait staggering and 
uncertain. On the 25th the animal had decidedly improved, and it 
was noticed that the oedema of the sheath and abdomen was 
distinctly less in amount; the animal seemed brighter and moved 
freely about the loose box. 

‘ This amelioration of the symptoms was gradually progressive, 
and on the 2nd July the oedema under the belly had completely 



4 


disappeared. On the 3rd July, however, the temperature began to 
rise steadily, and it continued high until the 9th, notwithstanding the 
fact that no haematozoa were present in the blood. During this 
time, the animal presented symptoms, which might be accounted for 
by the continued high temperature, although on the 6th symptoms 
of kidney disorder presented themselves, followed by epithelial casts 
and a small number of blood corpuscles in the urine. The large doses 
of Arsenic and Alkaloids have been persisted in, notwithstanding the 
intestinal and hepatic derangements, as it was feared that the high 
temperature indicated the probable return of the Surra haematozoon. 
On the 9th there were symptoms of gastro-intestinal catarrh, and 
taking these complications into consideration, as well as the fact that 
the haematozoa had been absent from the blood for a period of 
twenty days, it was decided to discontinue the Arsenic and Alkaloids 
entirely, and a laxative and simple febrifuge was substituted. 

4 From the 10th to the 14th all went well, and the animal seemed 
to improve, the temperature remained low, and the animal became 
much brighter and fed welL On the 15 th, however, he was dull, and 
the respiration and pulse had increased in frequency, the visible 
mucous membranes becoming of a dirty-yellow colour and 
petechiated. On the 16th, although his appearance was much 
brighter, the respiration had risen very high, and the heart’s action 
was irritable; the symptoms of gastro-intestinal catarrh and the 
kidney disorder had entirely disappeared. On the 17th the animal 
appeared much in the same condition, but the respirations were less 
although the temperature was a little higher. On this date one 
Surra haematozoon was discovered in two cover-glass specimens of 
blood, and this was the first organism observed during a period of 
twenty-eight days. The previous treatment of Arsenic and Cinchona 
alkaloids was at once resumed. On the 18th the animal had not 
altered much in appearance, the respirations were greatly improved 
in character and the pulse in frequency, but the temperature had 
risen. The blood contained a few haematozoa, but not so active in 
their movements as the one noticed the day before. On the 19th 
the condition was worse, the haematozoa very numerous in the blood, 
and the temperature much elevated, viz., 41 °C. On the 20th the 
appetite was normal, but otherwise the animal did not manifest any 
improvement; only two haematozoa were observed in two cover-glass 



5 


specimens of the blood, and these were exceedingly torpid On the 
2 ist the haematozoa were entirely absent, and from this date the 
animal appeared unable to throw off the effect which the return of 
the haematozoon had given rise to, and he gradually became weaker, 
and died on the 29th of July, although the haematozoa were absent 
during the last nine days. 

‘ It was decided in this case to try again the effect of the mixed 
Cinchona alkaloids and Arsenic on the haematozoon, as it had shown 
promise of efficacy in that of Horse No. LXV, in which case after a 
short exhibition of the drugs, the haematozoon was kept in subjection 
for a period of twenty-seven days. Consequently as a fresh trial was 
resolved upon, the animal was first put upon small doses, beginning 
with one drachm of the Alkaloids and four grains of Arsenic, given in 
the form of the Liq. arsenical is. On the morning following the first 
dose, the temperature was found to have fallen over a degree, and 
the haematozoon was absent from the blood, the temperature 
remained low, and the organism absent for the next five days, during 
which period two doses of the medicine were administered daily. 

' On reference to the Chart, it will be seen that on the ninth day 
of the disease the haematozoon again appeared in the blood, 
consequently on the tenth the doses of Alkaloids and Arsenic were 
increased to one and a half drachms and five grains respectively; 
these were continued for a period of eight days, during which time 
the haematozoon varied in numbers between few and very numerous. 
On the eighteenth day of the disease a further increase in the doses 
was agreed upon, and during the next four days they were put up to 
Alkaloids two and a half drachms and Arsenic grains vi, and for the 
latter half of this period the organism was absent from the blood, but 
re-appeared on the twenty-second day, and remained on the twenty- 
third at few. On the former date the Alkaloids were increased to 
three drachms, and the Arsenic to grains viii, and from the twenty- 
third to the thirty-fifth day of the disease, on which latter date the 
animal succumbed, the haematozoon was entirely absent from the 
blood. During the thirty-two days the animal was under treatment, 
three hundred and fifty-five grains of Arsenic in the form of Liquor 
arsenical is, and one hundred and fifty-four and a half drachms of the 
mixed Cinchona alkaloids were administered/ 

He further quotes the case of an animal belonging to the Bombay 



6 


Sodium arseniate 


Tramway Company. ‘ On the morning of the 7th October, 1892, the 
“ Surra” haematozoon was discovered on microscopical examination 
of its blood. On admission to the Laboratory Hospital the following 
note was made : — 1 “ Bay gelding, aged five years ; appears to be in 
perfect health, and in splendid condition.” On this date the 
haematozoon was absent from the blood, while for the next two days 
it was swarmingspecific treatment was deferred, a simple febrifuge 
draught being given to try and reduce the temperature. On the 
evening of the 12th October, the organism was only present in small 
numbers in the blood, and the temperature had decreased 
considerably, as had also the pulse. It was decided to again attempt 
the destruction of the haematozoon by the exhibition of Cinchona 
Alkaloids and Arsenic, commencing in this instance with large 
doses (viz., Alkaloid drs. iii, and Arsenic grains iv, bis die). 
The reason why large doses of the former were exhibited was 
that few haematozoa remained in the blood, and in order to 
decide whether their destruction could be compassed by the 
drug, the dose was so regulated as to ensure its having a fair 
trial. No marked change took place in the animal’s condition, nor in 
the number of the organisms present, until the 16th instant, when 
the haematozoa became numerous, the temperature rose considerably, 
respirations were increased in number, and it was noticed that the 
sheath was swollen ; the treatment was continued, the dose of Arsenic 
having been gradually raised to six grains. On the 17th the 
organisms had again fallen to few, but the temperature and pulse 
were still above normal. On this date it was decided to try the effect 
of full doses of Quinine , as this drug has been said to be of much 
value when exhibited in this manner. Ranking has stated that “ by 
the employment of full doses of Quinine and Arsenic he has been able 
to cure the disease.” It appeared from our previous experiments on 
the use of similar drugs, that it would be futile to attempt the 
destruction of the haematozoa by giving only the full medicinal dose, 
and we considered it to be advisable therefore to commence with a 
dose of drs. iv of quinine, and arsenic grains xii, in the 24 hours, 
carefully watching its effects, and in case of any untoward symptorti 
appearing, to discontinue its use.’ 

LAVERAN and MESNIL in 1902 published in the Annales de 
1 ’Institut Pasteur their researches ‘ Sur le Traitement et la prevention 



7 


du Nagana.’ They obtained their best results by using sodium 
arseniate subcutaneously; with this drug they were able to bring 
about a quick disappearance of the parasites from the blood. The 
lives of the animals were prolonged, but a permanent cure was not 
effected. These observers also used Sodium cacodylate , Sublimate 
of Mercury , various salts of Silver , but without effect 

They were also led to try the effects of injecting human serum Serum 
into ngana-infected rats and mice ; they were led to do so as man 
was not susceptible to ngana. A marked improvement followed the 
injection of human serum ; the duration of the disease was prolonged, 
but a real cure could not be effected. 

E. J. MOORE, in a paper upon the beneficial effects of Sodium 
arseniate employed hypodermically in tsetse-fiy disease, and published 
in the Lancet , Vol. II, 1904, records the most marked beneficial effects 
with large doses, and states that he would also recommend it for 
humans. 

CHICHESTER, who collaborated with him in a letter which he sent to 
Sir Patrick Manson dated May 5th, 1904, and published in the Lancet 
under the title of 4 Arsenic in the treatment of Trypanosomiasis in 
Cattle in Nigeria/ says that 4 he used Arsenic hypodermically and 
produced most wonderful effects.’ He adds that the experiment is 
not over, but says that f I do not think it wise to wait longer. I tell 
you what I have found, and you may perhaps think it wise to ask 
others to try the same treatment, especially in those parts where it 
seems a scourge to human beings.’ 

THOMAS, in 1903, whilst at the MacGill University, in Canada, 
repeated some of the experiments upon the action of Arsenic on 
ngana-infected animals, and on joining the Liverpool School of 
Tropical Medicine, in August of the same year, immediately started 
extensive investigations upon the action of drugs upon trypano¬ 
somiasis. Amongst them he used Sodium arseniate , and with this 
drug his results were similar to those of Laveran. 4 Many of the rats 
either died from the disease, were killed by the drug, or succumbed 
to an extensive ulceration around the site of injection.’ Thomas’s 
own words, taken from his paper entitled 4 Some experiments in the 
treatment of trypanosomiasis’ ( British Medical Journal , May 27, 

1905) were: 4 Arsenic in the treatment of trypanosomiasis in animals 
seems rather to mitigate the disease, to cause the parasite to 



8 


apparently disappear from the animal’s blood, and to prolong the life 
of the animal. Unfortunately the usual history is that if treatment 
be discontinued, even after a prolonged course of the drug, the animal, 
after a varying length of time, once more exhibits the symptoms, and 
finally succumbs to the disease. Undoubtedly some of the more 
resistant animals do recover.’ Thomas then quotes the view of 
Laveran and Mesnil (Trypanosomes et Trypanosomiases) that 
Arsenic kills the parasites which are free in the blood, but that when 
once the drug is eliminated or fixed in the tissues, the surviving 
parasites commence to multiply and the organisms once more 
reappear in the peripheral circulation unless another injection of 
Arsenic be given. On the administration of a second dose the 
parasites disappear only to reappear, and even though treatment be 
kept up, the majority of the animals succumb either from the disease 
or from the toxic effects of the drug. Some of the animals, such as 
rats and dogs, have been cured. Laveran also records the extensive 
necrosis and the pain which is apt to follow the administration of 
Arsenious acid. 

Thomas and Breinl, in a paper upon ‘ Trypanosomiasis 
and Sleeping Sickness,’ published as Memoir XVI, 1905, of 
the Liverpool School of Tropical Medicine, report fully upon the 
treatment. Sodium arseniate was found the most useful form of 
Arsenic, but the usual disadvantages soon appeared and they stopped 
it. The sum total, therefore, of results obtained with Arsenic or its 
salts, dating from the early observation of Livingstone, through the 
elaborate experiments of Bruce and Lingard, to the more recent 
experimental observations of Laveran, Mesnil, Thomas, and Thomas 
and Breinl was such as to encourage hope of a successful treatment; 
there was no doubt that the disease was modified, the life of many of 
the animals was prolonged, and some of the animals were cured. 
All experiments showed equally well the disadvantages of the drug, 
such as the recrudescence of the disease on stopping the drug and 
the severe toxic symptoms caused by it. 

Ehrlich and Shiga at about this time (1904) published their 
1 Farbentherapeutische Versuche bei Trypanosomenerkrankung' in 
the Berliner klinische Wochenschrift, 1904, Nos. 13, 14. These 
observers were the first to introduce colouring matters belonging to 
the Benzopurpurin group in the treatment of trypanosomiasis; they 



9 


obtained better results with Trypan-red than with either Arsenic or 
human serum, upon mice infected with Mai de caderas. They were 
able to cure mice, and in rats the parasites disappeared, but reappeared 
after a short time. The result of these eminent observers drawing 
attention to the use of Anilin colours was to stimulate a great amount 
of investigation in this direction. 

LAVERAN and MESNIL, ‘ Le trypanroth dans le traitement de 
quelques Trypanosomiases 1 (Comptes rendus de l’acad. des sciences, 
VoL 139, p. 19), confirm Ehrlich and Shiga. 

Halberstadter (Centralbl. f. Bakt., 1905, Vol. 38, p. 525) 
confirms Ehrlich as regards the action of Trypan-red in Caderas, 
similarly in Dourine, but had very little success in Ngana. 

NlSSLE (Arch. f. Hygiene, 1905, Vol. 53, p. 181) found Trypan-red 
better against Ngana than against Caderas. 

EWALD FRANKE, 4 Therapeutische Versuche bei Trypanosomen 
erkrankung’ (Inaugural dissertation, 1905, Jena), as the result of an 
extensive study of Trypan-red on T. equiperdum and Mbori strains, 
recommends especially the combined treatment of Trypan-red and 
Arsenic. 

WENDELSTADT (Deutsche med. Wochenschrift, 1904, No. 47) 
found that Trypan-red administered in small doses internally caused 
the trypanosomes to disappear, but that they soon reappeared again. 
He was able to cure rats infected with Ngana. 

WENDELSTADT (Deutsche med. Wochenschrift, 1907) by the 
internal application of trypan-red only succeeded in obtaining 
prolongation of life in the case of ngana-infected animals. 

THOMAS and Breinl in a paper entitled 4 Trypanosomes, 
Trypanosomiasis and Sleeping Sickness' (Memoir XVI, 1905, of the 
Liverpool School of Tropical Medicine), published a full report of 
treatment experiments with various drugs, amongst them Trypan-red , 
but they only succeeded in demonstrating that it helped to prolong 
the life of infected animals. 

WENDELSTADT and Fellmer, in a paper entitled 4 Uber die 
Einwirkung von Brilliantgrun auf Naganatrypanosomen ’ (Zeitschrift 
fur Hygiene und Infekt Krankheiten, Vol. LII, 1906, pp. 263-280), 
state that the parasites disappear from the peripheral circulation, 
and that after repeated doses the blood of the animals becomes 
negative as tested by subinoculations. 



IO 


MESNIL and NlCOLLE, ‘ Traitement des Trypanosomiases par les 
couleurs de benzidine ’ (Premiere partie, etude chimique. Seconde 
partie, etude experimentale. Annales de l’lnstitut Pasteur, Tome XX, 
June, July, 1906), tried to find out the connection between the 
chemical constitution and the trypanocidal action of the benzidine 
colours. P. di-amido-diphenyl urea + Ac. H. (Ph.) and p. di-chloro- 
benzidine + Ac. H. (Cl.) were the only colours which showed any 
marked action on the trypanosomes. Amongst the Arsenic 
preparations with which they experimented Atoxyl was found to be 
the most efficient. 

MESNIL, NlCOLLE and Aubert (Annales de l’lnstitut Pasteur, 
January, 1907) give their experiments upon, amongst other drugs, 
the benzidine colours , and state that the blue colours are superior to 
the red colours, their best results being obtained with p. di-amido- 
diphenyl urea + Ac. H. (Ph.). ♦ 

MESNIL and NlCOLLE (Annales de l’lnstitut Pasteur, December, 
1907) describe in a third paper the final results of their experiments 
in the case of twelve monkeys infected with T. gambiense , of which 
six were cured by Atoxyl alone, four by the combined treatment of 
Atoxyl and Ph., and two by Ph., first employed alone, followed up by 
only one injection of Atoxyl in the first case, and two in the second. 

EHRLICH, in a paper entitled ‘ Chemotherapeutische Trypano- 
somenstudien ’ (Berl. klin. Wochenschrift, Nos. 9-12, 1907), says 
Benzidine colours (Trypan-red), T riphenylmethane colours {Brilliant 
green and Malachite green , Wendelstadt and Fellmer) have been 
found to be trypanocidal. Ehrlich experimented with Parafuchsin 
by feeding mice on parafuchsin cakes, which gave very good results. 

WEBER and Krause, in a paper entitled ‘ Farbstoffbehandlung 
der kiinstlichen Trypanosomeninfektion ’ (Berliner klinische Wochen¬ 
schrift No. 7, 1907), tested systematically different colouring matters 
(Crystal violet , Victoria blue , Fuchsin), as regards their trypanocidal 
action in Ngana, with the view to find relations between chemical 
constitution and action. Fuchsin seemed to have the best effect, also 
because of its relative harmlessness for animals. They were unable 
tb obtain cures, but the advantage of Fuchsin seems to reside in its 
power of prevention. 

WENYON, in a paper entitled ‘ Action of the colours of Benzidine 
on mice infected with Trypanosoma dimorphon ’ (Journal of Hygiene, 



II 


Vol VII, April, 1907}, describes how he treated mice infected with 
T. dimorphon and 54 Benzidine colours. He finds, in contradiction 
to Mesnil, Nicolle and Aubert, that the red colours have a more 
powerful effect on T. dimorphon than the blue ones. 

PLIMMER and Thomson have used Mesnil’s, Nicolle’s and 
Aubert’s Cl. colour (J>. di-chloro-benzidine + Ac. H .) and obtained the 
same results as with Trypan-red. 

KOCH, in a paper entitled 4 Schlussbericht ii. d. Thatigkeit d. 
deutschen Expedition z. Erforschung der Schlafkrankheit' (Deut. 
med. Wochenschrift, No. 46, 1907), states that Afridol blue and 
Afridol violet had not the least effect upon trypanosomes, neither 
had para-Fuchsin nor par a-Rose aniline. 

C. Browning, ‘ Experimental Chemotherapy in Trypanosome 
infections * (Brit. Med. Jour., No. 2,446, November 16th, 1907, p. 1405), 
gives the results of colour treatment, especially with para-Fuchsin. 
He recommends treatment with Atoxyl and Dye. 

YAKIMOFF, ‘Zur Behandlung der Dourine’ (Centralbl. f. Bakt. 
Orig., VoL XLV, h. 5, Dec. ’07) used Trypan-red for treatment of 
Dourine in mice, rats and rabbits. Several injections of Trypan-red 
are able to prevent relapses in mice and to effect a cure. During the 
incubation period, if Trypan-red is given very early, the appearance 
of trypanosomes is prevented. 

It does not act prophylactically, and there is no immunity after 
cure. The mechanism of Trypan-red action is tiypanolytic, 
trypanosomes being killed by Trypan-red by immune substances and 
other products. The immunity, however, is of very short standing. 

THOMAS in his paper referred to above, and published May, 1905, 
states that after the publication of Ehrlich's and Shiga’s results with 
Trypan-red, he repeated the experiments with animals infected with 
different species of trypanosomes^ ‘ The best results were obtained 
with mal de Caderas-infected animals; the results were not so good 
with animals infected with nagana and surra, and still worse in the 
case of animals infected with dourine, the Gambian horse strain, and 
T. gambiense. The parasites disappear for a few days to reappear, 
and the duration of the disease was not greatly prolonged, and on 
analysing the evidence given above of those who have carefully 
experimented with the aniline dyes, we are driven to conclude that 
the colours do not possess any advantage over the arsenic salts, that 



12 


they are not even as efficacious.' In other words, sodium arseniate 

had given better results and still held out to investigators a more 

promising field for ultimate success than the use of colours. 

Use of the com- However, following up the experiments of Laveran, Thomas 
bined treatment of 

Arsenic and colours determined to try a combination of the two drugs Trypan-red and 
Arsenic and the results were more encouraging, but he states, 
‘ unfortunately trypan-red also caused a nephritis, and by its chemi- 
otaxic properties very extensive necrosis sometimes resulted. On 
monkeys, especially, the subcutaneous injection of the dye either 
alone or in combination with arsenious acid induced ulceration, which 
so undermined the health of the animals that they succumbed to any 
outbreak of disease, which occurred only too frequently amongst my 
animals. It was these untoward accidents which induced me to seek 
a preparation of arsenic less toxic and the subcutaneous injection of 
which entailed less danger of necrosis .’ Before, however, proceeding 
to describe the compound of arsenic ( Atoxyl ), which he demonstrated 
had such a marked action upon the trypanosome, we wish to record 
the observations made upon the combination of Arsenic with colours 
and other bodies. 

THOMAS and Breinl, in their publication referred to above, 
came to the conclusion:— 

1 That in trypan-red we possess an agent of some use in the 
treatment of trypanosomiasis. That certain trypanosomic diseases 
appear to be more amenable to its action than others. That in the 
substance at present available there is need for improvement in order 
to abolish its toxic effects. 

1 That a combination of arsenic and of an improved form of 
trypan-red would seem indicated in the further investigation of the 
cure of trypanosomiasis.’ 

Laveran wrote a paper entitled 1 Traitement mixte des Trypano¬ 
somiases par l’acide arsenieux et le trypanroth des infections au 
Trypanosoma gambiense' published 30th January, 1905, in the 
Comptes rendus de l’Academie des Sciences. He was the first to 
use a combination of trypan-red and arsenious acid. He made a 
large number of experiments upon this combination, and we owe to 
him most of our knowledge upon the effects of this combined treat¬ 
ment upon small animals. He demonstrated the curative action of 
the combination of trypan-red and arsenious acid in the case of rats 





and mice infected with Mbori or Surra. In the case of animals 
infected with T. gambiense the results appeared to him less 
encouraging. 

In a second paper upon the mixed treatment, published April 
17th, 1905, in the Comptes rendus de TAcademie des Sciences he 
discusses the combined treatment of monkeys infected with 
T. gambiense. His observations confirm the results previously 
obtained with other animals. We reproduce his own comments upon 
the combined treatment in the case of T. gambiense. 

* II n*y a pas de motif pour que le traitement qui a reussi dans les 
infections experimentales du rat, du chien et des singes par 
Trypanosoma gambiense ne reussisse pas egalement dans les infec¬ 
tions naturelles, chez l’homme, et je crois que, des maintenant on 
serait autorise k essayer de ce traitement chez les sujets atteints de 
trypanosomiase. La difficult^ sera de determiner les doses d*acides 
arsenieux et de trypanroth qui devront etre preserves; des tatonne- 
ments seront inevitables. Les chances de succes seront d’autant 
plus grandes que la maladie sera a une p6riode moins avancee de son 
evolution. II est douteux que le traitement puisse donner encore de 
bons resultats quand les accidents du cote du systeme cerebrospinal 
ont acquis une certaine intensite: on se rappellera d’autre part que 
le trypanroth est irritant pour les reins (1), on surveillera les urines 
et Ton ne prescrira pas ce medicament aux malades atteints de 
nephrite.* 

FRANKE (loc. cit.) also strongly recommended, as the result of his 
extensive trials, the use of the Trypan-red-arsenic treatment. 
WENDELSTADT and FELLMER also advocate the treatment with a 
combination of Arsenic and Brilliant green. They also combine 
Brilliant green and Nucleinic acid. 

MAGALHAES, in a paper entitled 1 De 1 * Action des Composes 
arsenicaux et du vert brilliant sur le Tryp. gambiense et le Tryp. 
brucei * (Arch. de. R. Inst. bact. Camera pestana, T. I, Jan. 2nd, 1907), 
treated rats infected with T. gambiense with Sodium arseniate and 
Brilliant green. The parasites disappeared for a time but reappeared 
again. 

As seen from the above notes, the combined Arsenic-colour 
treatment led Laveran to believe that as it had succeeded in his hands 



x 4 


so well in the case of animals artificially inoculated with T. gatnbiense , 
so it might succeed in man in the case of sleeping sickness, 
^oxyf^try^no- Thomas, however, as we have seen, had given it up on account 
somiasis in animals of the nephritis and local necrosis which was induced, and set himself 
to find a preparation of Arsenic less toxic than Arsenious acid or 
Sodium arseniate, and the subcutaneous injection of which produced 
less danger of necrosis. An aniline compound, the anilide of met- 
arsenious acid (Atoxyl), having the supposed formula C fl H 5 NH AsO a , 
attracted his attention. This preparation had been before the medical 
profession since 1900, and various workers had recorded its use in 
the treatment of various skin affections and in anaemia. 

F. BLUMENTHAL, in a paper ' Uber Metaarsensaure anilid * (Die 
medizinische Woche., 14th April, 1902, No. 15), describes how he had 
used Atoxyl on rabbits in order to test the toxicity of the drug. He 
concluded from his experiments that Atoxyl was 40 times less toxic 
than Solutio Fowleri. 

SCHILD (Berl. klin. Wochenschrift, 1902, No. 23, and Dermato- 
logische Zeitschrift, Band X, Heft 1, 1903) employed Atoxyl against 
different skin diseases as psoriasis, lichen ruber, etc, with very good 
results. 


F. BlRINGER published in the Therapeutische Monatshefte, 
August, 1903, a paper on ‘ Klinische Erfahrungen mit Atoxyl. * He 
used Atoxyl in the treatment of different skin diseases, and as a 
result regarded it as a valuable and welcome substitute for Arsenious 
acid. 

MOLLER, in a paper in the Berliner klin. therap. Wochenschrift, 
entitled f Uber die Heilung der Tuberkulose mit Atoxyl/ employed 
Atoxyl administered intravenously in 50 cases of tuberculosis with 
very encouraging results. 

F. Mendel (Therapeutische Monatshefte, April, 1903), * Zur 
endovenosen Application der Medikamente/ states that Atoxyl is 
especially suitable for either subcutaneous or intravenous administra¬ 
tion. It produces no necrosis, no fever, and very much larger doses 
of arsenic can be given without producing toxic results. ‘ I have 
myself / wrote Thomas, ‘ tried the drug in high doses intravenously 
zuithout ill effects! 

To Thomas, and to Thomas and Breinl, belongs the credit of 
having, after a very careful experimental investigation, introduced 



15 


to the notice of the medical profession the superior claims of Atoxyl 
as a curative agent in cases of trypanosomiasis. Before, however, 
recommending it to the profession, they put it to a most searching 
test in animals. We quote Thomas’s own words:—‘Tentative 
experiments on rats and rabbits appeared so favourable that I 
decided to institute a series of experiments. These observations 
have been in progress for the last ten months, and have on the whole 
been most promising. In all my therapeutic experiments certain 
conditions were laid down to be followed out. 

1. That the animal should be well infected and the presence of 
parasites determinable in its blood. 

2. That the disease should have been established some time 
(one cannot expect to treat either man or beast suffering from the 
disease in the very early stages when no definite symptoms are 
manifested). 

3. That some symptoms besides the presence of parasites should 
be in evidence: 

{a) Anemia; 

(b) Loss of weight. 

‘ If the treatment of such infected laboratory animals be successful, 
then such a line of medication ought to be of service in the practical 
treatment of the disease. 


Trypanosomes 

The trypanosomes experimented with are the following : — 
Trypanosoma gambiense — 

(a) Gambian fever strain (Gunjur). 

(b) Congo Free State fever strain. 

(c) Uganda sleeping sickness. 

( d ) Congo Free State sleeping sickness. 

(e) A highly virulent strain derived from one of my cases of 

sleeping sickness which had only been passed through a 
monkey, baboon, and a rabbit. 

Trypanosoma brucei (nagana). 

Trypanosoma evansi (surra). 

Trypanosoma equinum (mal de Caderas). 

Trypanosoma equiperdum (dourine). 

Trypanosoma dimorphum (Gambian horse disease). 



i6 


Animals Used 

T. gambiensc Monkeys, dogs, pups, kittens, rabbits, guinea-pigs, 
rats and mice. 

T. evansi. —Rabbits, guinea-pigs, dogs, rats, mice. 

T. brucei .—Rabbits, guinea-pigs, rats, mice. 

T. equinum .—Dogs, rabbits, guinea-pigs, rats and mice. 

T. equiperdum .—Pups. 

T. dimorphum .—Dogs, pups, rabbits, guinea-pigs, rats and mice/ 

We reproduce also his conclusions upon the advantages of Atoxyl 
in trypanosomiasis:— 

4 With five exceptions every animal has had one or more controls 
which were infected at the same time ; the controls of T. gambiense , 
strain 4 e' surra, nagana, caderas, and Gambian horse parasites have 
all died in the usual time. It is, therefore, evident, from the great 
majority of the experiments, that the treatment of animals infected 
with trypanosomes with this preparation or in combination with the 
dye either arrests the disease, thus prolonging the life of the animals, 
or apparently cures them. This is especially the case in animals 
infected with the ordinary strain of T. gambiense , and even, though 
to a less extent, with the abnormally virulent strain called ‘ e .* 

1 A comparison of animals infected with the same strains, but 
treated according to Laveran's method with sodium arseniate has 
made me conclude that treatment with this aniline compound is in 
many ways superior to the ordinary arsenical treatment, on account 
of the quicker action of the drug on the parasite, the fact that the 
action seems to be prolonged, that large doses can be given without 
toxic symptoms and the entire absence from any tendency to cause 
sloughing. 

4 In my opinion treatment is indicated, of cases of trypanosomiasis 
in man, with this drug in high doses administered intravenously and 
for a long period, pushing it to the maximal amount that the case can 
stand without headache and nausea, at the same time building up 
the patient in every way possible that will conduce to a lessening of 
the anaemia. In the European the combination of trypan-red would 
probably be objectionable on account of the intense colouring of the 
tissues and secretions, but the native exhibits only a reddening of the 
conjunctivae and staining of the secretions. One of my cases of 



l 7 


trypanosome fever had been under treatment with trypan-red and 
arsenic for a short time before he returned to the Congo with favour¬ 
able results—the parasites were longer absent from the peripheral 
circulation ; his general blood condition was better; his temperature 
was almost normal. 

‘ I present the results of my experiments tentatively. I have used 
the term “ apparently cured,” as any one with an intimate knowledge 
of T. gambiense and other forms of trypanosomes in animals knows 
how difficult it is to say that an animal is not infected. This 
is especially the case with the human parasite. In a previous 
paper I have recorded cases in which the blood has been negative for 
nearly a year in rats known to have been infected, and which at the 
expiration of that time showed parasites once more in their blood. 

* I do not believe that sodium arseniate alone will be found of 
great practical value, nor do I think atoxyl is a perfect preparation, 
from its toxic effects on canines and felines, but it is an advance on 
arsenious acid, and, if further efforts be made to produce a substance 
like trypan-red, but less irritating in action, the combination ought 
to be of service in the treatment of trypanosomiasis in man.’ 

THOMAS at about- the same time made a communication to the 
Royal Society (received April 8th, 1905, and read May nth, 1905) 
upon the experimental treatment of Trypanosomiasis in animals with 
Atoxyl, Atoxyl and Trypan-red, and Trypan-red alone. The com¬ 
munication was the result of exhaustive experiments upon animals 
infected with five strains of T. gambiense , one a very virulent one 
taken from a case of sleeping sickness, and the rest the common 
animal strains. The Atoxyl was given in two ways. 

(1) High doses at intervals of a week. 

(2) High initial dose and then reduced amounts administered 
three times a week. 

He concluded, ‘/n my hands the arsenic-anilin compound ( Atoxyl ) 
has given far better results than treatment with sodium arseniate. The 
advantages of its administration intravenously or subcutaneously in high 
doses over a length of time — namely , its less toxic properties , the absence of 
all tendency to cause sloughing and the apparently longer action of the 
drug , make me believe that the employment of this compound is indicated 
in the treatment of human trypanosomiasis * 

A third paper by THOMAS in collaboration with BREINL upon 
c 



i8 

‘ Trypanosomes, Trypanosomiasis and Sleeping Sickness,’ was pub¬ 
lished as Memoir XVI, 1905, of the Liverpool School of Tropical 
Medicine. 

This paper contains the full report of all the treatment experi¬ 
ments. They wrote: — 

‘ We have, therefore, to realise that the ordinary arsenic 
compounds when administered, only produce a temporary favourable 
effect, that, if long continued, the animals will die either from the 
parasite or from the arsenic, or from both. Hence, some other 
compound is indicated. It is for this reason that the newer 
compounds of arsenic have been experimented with in order to find a 
preparation capable of being used over a long period and in high 
doses without producing toxic symptoms Of the various preparations 
tried, a meta-arsenic anilin compound, atoxyl, has proved the most 
satisfactory, but it is not ideal. It is not non-toxic, as dogs, kittens, 
guinea-pigs, and rabbits have shewn toxic symptoms and succumbed, 
but it is not so toxic as sodium arseniate. It does not produce the 
sloughing which so often follows the subcutaneous or intravenous 
inoculation of sodium arseniate, it causes no pain, and its administra¬ 
tion can be continued over a period of many months even when used 
in extremely high doses. 

4 It is the only remedy at present giving any prospects of a cure. 
In the treatment of cases a rational method of treatment must be 
adopted. It is useless, for instance, to only administer arsenic for a 
short period or until the parasites have apparently disappeared from 
the peripheral circulation. The drug must be administered in as 
high doses as possible, and it must be continued even until after all 
the favourable signs are present, viz., disappearance of parasites from 
the blood, increase of weight, improvement in the blood count 
and percentage of haemoglobin, loss of the auto-agglutination 
phenomenon of the blood corpuscles, decrease in and more regular 
temperature. From time to time susceptible animals ought to be 
inoculated with large quantities of the patient’s blood, at least in 
amounts of 5*0 to 15’oc.c. At the same time all aids in building up 
the physical condition of the individual should be used If such a 
regime be carried out, and treatment commenced at an early period, 
the prognosis (based on the experience of treated animals) will be 
good. 



*9 


‘ The drug was used only upon animals showing the effects of the 
parasites, such as loss of weight, anaemia, fever, and auto-agglutination 
of the corpuscles, and no animal was used until its blood contained 
numerous parasites. The numbers of the parasites present differed 
according to the species of animal and the disease. In the majority 
of the experiments, control animals which were not treated, and had 
been inoculated at the same time as the treated animals, were used. 
In all cases the control animal died. 

1 Intravenous inoculation was used only on rabbits, all other animals 
were injected subcutaneously. Treatment was continued for one to 
three months or until increase of weight, diminution of the anaemia, 
and entire absence of parasites from the blood, as far as microscopical 
examination could determine, was noted. At various periods 
susceptible animals were inoculated with the blood from a treated 
animal. When treatment had been discontinued for one to three 
months, or longer, the animal was bled or killed and all the blood 
available was used to inject susceptible animals. Inoculated animals 
whose blood has given negative results after three to six months, or 
after longer periods, have been inoculated with virulent blood, and 
have taken the disease, thereby showing that no immunity was 
conferred by the previous inoculation. 

‘ T. gambiense. —Rabbit, male, weight, 2,010 grammes. Parasites 
appeared on the twelfth day. On the forty-sixth day numerous 
trypanosomes were present; it had lost weight (1,890 grammes). A 
blood count gave reds, 4,980,000; whites, 8,860; haemoglobin, sixty- 
seven per cent. For three-and-a-quarter months it received ro c.c. of 
five per cent, solution atoxyl three times a week, gradually increasing 
the amount to roc.c. of ten per cent, solution. It then weighed 
2,000 grammes. The blood count was reds, 6,640,000 ; whites, 6,200 ; 
haemoglobin, eighty-eight per cent. The blood in quantities of 10 c.c. 
was non-effective. The auto-agglutination of the corpuscles was lost. 
Thirty-two days later it was very ill; it was therefore bled to death, 
and the whole of its blood injected into a monkey. This monkey 
has never become infected. The post-mortem showed severe 
haemorrhagic cystitis, the bladder in parts being almost gangrenous 
and acute septic peritonitis, especially around the bladder. The 
spleen showed no congestion, but the connective tissue was slightly 
increased. The kidneys and liver were normal. 



20 


‘Rabbit, 889, inoculated October 26: weight, 1,760 grammes. 
Blood count: reds, 6,620,000; whites, 6,700; haemoglobin, eighty- 
nine per cent. Parasites were seen from November 8 up to January 
10; the trypanosomes were always present but in small numbers; 
they then increased to eighteen to twenty to a field. The anaemia 
was pronounced, and loss of weight was noted. It then weighed 
1,540 grammes. Blood count : red, 3,880,000; white, 11,800; haemo¬ 
globin, sixty-three per cent. It could hardly sit up, and remained 
most of the time lying down. This animal was given o’8cc. of five 
per cent, solution atoxyl. At the end of eighteen hours the parasites 
were absent from the blood. Doses were given twice a week, 
beginning with O'5 c.c., and increasing to 2*0 c.c. of a five per cent, 
solutioa The blood in large doses is non-infective. The animal is 
vivacious; the coat is smooth and thick. Average weight, 1,600 
grammes. Several guinea-pigs infected for about two months, and 
showing twenty to forty parasites to a field, have been treated. They 
were injected subcutaneously with 0*3 c.c. of ten per cent, solution. 
At the end of the fifteenth hour no parasites were seen. Treatment 
was o* 1 to 0 3 c.c. of five per cent, solution three times a week for two 
months. The animals all increased in weight. One died sixty-two 
days after treatment was discontinued. Three rats inoculated with 
its blood never became infected. The second and third pigs were 
killed at the end of eighty and one hundred days after stopping 
treatment, and the blood used to inoculate controls. No control has 
shown the parasites. 

‘Virulent strain.—Rhesus.—Weight, 2,815 grammes. Inoculated 
November 16. Blood count: red, 5,220,000; white, 12,800; haemo¬ 
globin seventy-eight per cent. On November 31, parasites were 
seen. Two days later, twelve to seventeen to a field were noted 
The weight was 2,420 grammes. Blood count: red, 4,600,000; 
white, 7,200; haemoglobin, seventy-three per cent. The parasites 
counted per mm. 3 gave 100,000. Oedema of the eyelids and bridge 
of nose was present The animal was given o*8 c.c. of ten per cent, 
solution atoxyl subcutaneously. Four hours later: red, 4,450,000; 
white, 24,800; parasites 40,000 per mm. 3 At the eighth hour: red, 
4,740,000; white, 25,200; parasites one to eighty-nine fields. 
Between the fourth and eighth hours the parasites were seen to 
become remarkably degenerated and deformed ; many phagocytes 



were present. At the twenty-fourth hour after injection a count 
gave red, 5,050,000; white, 46,000. The blood was negative. The 
leucocytes remained high for a couple of days and then fell; in none 
of the phagocytes could any remains of trypanosomes be found. 
Treatment: rocc. of ten per cent, solution was given twice a week. 
The animal increased in weight. The auto-agglutination of the 
corpuscles began to be less accentuated, and the number of erythro¬ 
cytes and the haemoglobin rose. The local oedema disappeared 
On the thirty-ninth day dysentery appeared, and the animal 
succumbed on the forty-eighth day after injection. The autopsy 
showed a very severe haemorrhagic and necrotic enteritis, with 
slightly enlarged spleen. Kidneys normal. Glands, small; inguinal 
group haemorrhagic. The blood was non-infective in amounts of 
ro cc., but infective if 15 c.c. of pure blood was used. Unfortunately, 
the arsenic was discontinued on the appearance of dysentery. A 
second monkey, inoculated from the first Rhesus just before treat¬ 
ment was begun, was treated with the same doses of arsenic; the 
parasites disappeared in the same way, but the animal quickly 
succumbed to dysentery. 

‘ Many rabbits inoculated with this strain have been treated. It 
was found that unless treatment was started early that the majority 
of animals died as it was so exceedingly virulent. With these animals 
treatment was begun earlier and higher doses given than with the 
standard “ Gunjur ” strain. Despite treating the animals early, some 
died. With this strain treatment had to be kept up longer. Some 
rabbits have survived eight months after injection, while all the 
controls have died in fourteen to thirty-six days. Guinea pigs 
infected with this strain do not react so well to the treatment. Rats 
must be treated early and with high doses if treatment is to be 
successful. Mice infected with this strain react if treatment is 
commenced early enough. The action of atoxyl on the various 
trypanosomes has been studied, and after numerous observations, 
continued for the whole period during which the drug was adminis¬ 
tered, the effect appears to be as follows: — 

‘ On administration of arsenic compounds into an animal showing 
numerous parasites in the blood the following action on the trypano¬ 
somes will be noticed. For the first three-and-a-half to four hours, 
depending on the dose used, very little change in the parasites can 



22 


be noticed. Between the fourth and fifth hour the effect on the 
trypanosomes is evident Some parasites appear to be swollen and 
their movement is less rapid. If now a series of blood specimens be 
examined at intervals of twenty to thirty minutes, the following 
changes will be seen. The number of slowly moving trypanosomes 
increases, many parasites will be seen to be almost motionless. The 
protoplasm takes on a peculiar ground-glass appearance, and dark 
granules appear in the protoplasm; very often a small series of 
granules one behind the other, sometimes in pairs or all clumped 
together, are seen lying between the macronucleus and the anterior 
end, or distributed through the whole body of the parasite. At the 
same time vacuoles are observed, oftentimes very large. The 
trypanosomes become deformed, assuming various shapes, the most 
common being a kite-shaped form with fairly long flagellum, and a 
tadpole-like one with hardly any free flagellum. These forms 
especially exhibit greatly impaired movements. At the same time 
a noticeable increase of the leucocytes is discernible; phagocytes 
begin to appear, very often groups of five to seven will be seen. 
Up to this time (sixth to seventh hours) the trypanosomes, though 
decreased in numbers are still present in considerable quantities. 
Suddenly, in the course of an hour, the numbers may drop from 
forty to two to three to a field or less; coincident with this is a 
very marked increase in the number of leucocytes, especially phago¬ 
cytes. From the ninth to the fourteenth and sixteenth hours the 
changes are less pronounced and rapid, the trypanosomes gradually 
disappear. At the eighteenth hour, provided the animal has been 
injected with the correct amount the parasites are absent from the 
peripheral circulation and, even though the blood is centrifuged, 
none can be found. 

‘ From a series of these observations, we have determined that in 
hardly any of the forty-six continuously observed animals were 
parasites to be found after the eighteenth. Should, however, the 
drug be given in smaller amounts the process takes longer, lasting 
from thirty-six to forty-eight hours. 

* From the experimental work with various therapeutic agents the 
following conclusions can be made : — 

‘(i)That animals suffering from trypanosome infection react 



favourably to only a few agents, of which arsenic is the only drug 
which seems to exert a more than transient action. 

4 (2) That the greater the amount of arsenic introduced into the 
system of the animal the greater and more permanent the effect on 
the parasite. 

4 (3) That arsenic medication is indicated in the treatment of 
individuals suffering from trypanosomiasis. That the treatment 
ought to be long continued and regularly administered in as high 
doses as the case can stand. That all aids to building up the system 
should be employed.’ 

In a fourth paper upon 4 Atoxyl in the treatment of Trypanoso¬ 
miasis,’ published in the British Medical Journal, Jan. 19th, 1907, by 
Drs. Breinl and Todd, these observers summarised our knowledge 
concerning the use of Atoxyl up to January, 1907. They quote Van 
Campenhout’s private communication, in which he refers to the 
combination of Atoxyl and Strychnine and a cold bath, the latter for 
a tonic and stimulating effect. Three Europeans treated by Van 
Campenhout have gained weight and are apparently well. He has 
obtained good results by the treatment of Europeans by Atoxyl, in 
the first stages of trypanosomiasis. He prefers a solution of 5 per 
cent, rather than 10 per cent. 

Todd and Breinl recommend the use of a 20 per cent, solution 
administered in increasing doses, up to 0*2 gramme. 

The following letter was published by Professor Ross to make it 
clear that atoxyl was first suggested and used by Thomas and Breinl 
in trypanosomiasis and sleeping sickness in man, as fully established 
in the preceding pages. 


THE TREATMENT OF TRYPANOSOMIASIS. 

Sir,—Many statements having been made recently in the lay press 
to the effect that trypanosomiasis has been cured by various persons 
by the means of atoxyl, I should like to point out that this drug was 
first suggested and used by Drs. Thomas and Breinl, of this School, 
for the purpose referred to. A full account of their experiments was 
given in the Proceedings of the Royal Society, November 9th, 1905, 
vol. 76, and also in our publications, Memoir XVI. At the instance 
of this School large quantities of the drug have been sent to the 



24 


Congo, and several patients are now under treatment with it. The 
result has been an apparent success so far, but it is of course too early 
to speak definitely as to the final result, because it is well known that 
patients may survive for several years, even without treatment, and 
yet ultimately succumb. For example, we have had cases under 
observation for four years without a fatal result.—I am, etc., 

RONALD ROSS, 

Professor of Tropical Medicine, 
Incorporated School of Tropical Medicine. 

Sept. 2 ist., 1906. 

Editor * British Medical Journal.* 

At the end of 1907 overwhelming clinical proof derived from the 
human subject is forthcoming, as we shall see. The statement that 
‘ We have in Atoxyl the specific drug for trypanosomiasis , as we have in 
Quinine that for Malaria,' fairly represents the view of Professor Koch. 

Sir Patrick Manson, who has had the largest experience of this 
drug in human cases of trypanosomiasis in this country, concludes in 
the following words : —‘ The prospects of atoxyl treatment I consider 
most hopeful. As regards efficiency and mode of action, it seems to 
me that it is almost on a par with mercury in syphilis and quinine in 
malaria; and I think in using atoxyl we should conform our practice 
to what experience has taught us to be the best methods of using 
these other efficient and long-tried remedies. I don’t believe we can 
kill the trypanosomes outright by one or two large doses of atoxyl, 
any more than we can kill the treponema of syphilis or the parasites 
of malaria by large doses of their respective specifics. Mercury does 
not immediately cure syphilis nor does quinine immediately cure 
malaria; buf they deprive their respective parasites of their patho¬ 
genic properties and keep the patient alive and in good health till, in 
process of time, the parasites either die out or become permanently 
inert.' 

By far the largest clinical experience of the drug has, however, 
been obtained by Professor Koch and his assistants, and their 
conviction is overwhelming on the advantages of this drug over all 
previous ones. 

R. KOCH, ‘ Bericht fiber die Tathigkeit der deutschen Expedition 
zur Erforschung der Schlafkrankheit bis zum 25 November, 1906* 
(Deutsche, med. Wochenschift. Jahr. XXXIII, No. 2, 1907). 



25 


Koch treated 986 cases of sleeping sickness with Atoxyl* Out 
of 336 cases positive results were obtained in 34.7. He points out the 
importance of the drug in early cases when it is much more efficient 
than in advanced cases. Koch divides his cases into two classes— 
‘ early * and * advanced * cases. He gives a short clinical account of 
the symptoms. Debility, nervous excitement, trembling of the 
extremities, general disturbance of the psychical functions, and 
nervousness very often in the form of mania. In the latest stages 
of this disease, apathy and sleep. Atoxyl was usually administered 
in the forms of two double injections of 0*5 gramme. Usually, after 
the second injection the parasites had disappeared from the blood 
and from the lymph glands, but a real improvement is only to be seen 
three or four weeks after the injection. In advanced cases, it was 
impossible to drive out the parasites from the circulation, even after 
prolonged administratioa Under the influence of Atoxyl, the trypano¬ 
somes sometimes disappeared for 30 to 40 days; when they 
reappeared they were only observed in very small numbers. Attempts 
to shorten the treatment by the administration of one large dose of 
Atoxyl only have given uncertain results as yet. In 12 cases, in 
which only one full dose of Atoxyl was given, the trypanosomes 
disappeared for 30 to 40 days. 

R. KOCH, 1 Schlussbericht iiber die Tatigkeit der deutschen 
Expedition zur Erforschung der Schlafkrankheit ’ (Deutsch. med. 
Wochenschrift, 1907, No. 46), gives his further experiences. 
He believes that some of the appearances, after Atoxyl 
treatment, can only be explained by the resorbtion of the dead 
trypanosomes. He still uses as the mode of administration the double 
injection of 0‘5 gramme Atoxyl on two successive days, then 10 days 
interval, and two more injections. Koch was not able to observe, 
as Erlich did, that the trypanosomes become resistant to the Atoxyl. 
Attempts to administer Atoxyl in doses of one gramme were 
unsuccessful in so far as after such large doses of atoxyl blindness 
very frequently occurred, due to atrophy of the optic nerve. Very 
interesting from an epidemiological point of view is the fact, which 
he has pointed out in his previous reports, that in Kisiba fifteen 
married women were found suffering from sleeping sickness. As 


Up to date he has treated some 2,000 cases. 



these women had never left the place, and it was found that their 
husbands were all either suffering from sleeping sickness or had died 
of that disease, Koch brings forth his own explanation: that 
trypanosomes may be transmitted by coitus. 

Koch quotes a further case, where three women were all infected 
with trypanosomes from one man. 

Koch states that early cases of trypanosomiasis can be definitely 
cured by Atoxyl by a six months treatment; in far advanced cases 
it is very difficult to drive out the parasites. 

BRODEN, in a paper entitled 4 Les Trypanosomes dans l’Etat du 
Congo ’ (Rapport sur les traveaux du laboratoire medicale de 
Leopoldville de 1900 a 1905, II, Bruxelles, Hayem, 1906, p. 71-143), 
used Atoxyl in the treatment of one case of trypanosomiasis with 
very encouraging results. 

BRODEN ET Rodhain ‘ Le traitement de la trypanosomiase 
humaine (Maladie du Sommeil)* (Archiv fur Schiffs und Tropen- 
hygiene, Band X, 1906, p. 693), treated cases of trypanosomiasis 
(three white men) with Atoxyl. Fever disappeared in all three and 
their general heath improved. 

Van CAMPENHOUT starts with o*2 gramme of Atoxyl, and 
increases his doses 0*05 gramme every second day till he reaches 
0*8 gramme. This does is given every second day for a fortnight or 
three weeks, and then he decreases his doses gradually 0*05 gramme. 
In the cases of the second period he uses strychnine and cold baths. 

Mesnil, Nicolle ET Aubert, 4 Recherches sur le traitement des 
infections experimentales a Trypanosoma gambiense ’ (Annales de 
l’institut Pasteur, Tome XXI, Jan., 1907, pp. 1 -19), continue the 
researches on a number of Benzidine colours and Sodium arseniate 
and Atoxyl on Trypanosoma gambiense. Amongst the Benzidine 
colours in general, the blue colours show themselves superior to red 
colours. The best results were obtained with p. di-amido-di-phenyl- 
urea 4- AcH (Ph.). Atoxyl is considered superior to Sodium arseniate. 
Further, they combined the colour Ph. with Atoxyl with very good 
results. 

KOPKE publishes his experiences in the following papers: 

4 Trypanosomiases humaines/ (XV Congres international de 
medecine de Lisbonne). 4 Traitement de la maladie du Sommeil,’ 
(Travaux de r£cole de Medecine Tropicale de Lisbonne). 4 Traite- 



2 7 


ment de la maladie du Sommeil/ (Rapport presente au XIV Congres 
Intern. d’Hygiene et Demographie. Lisbonne, 1907). 

His results with atoxyl are not very good Out of 29 cases, 22 
died from trypanosomes. Seven are still alive and of these only 
two are in good health. 

Kopke used very large doses of Atoxyl (one gramme in one 
injection). Out of 29 cases, six had eye lesions which were 
diagnosed as atrophy of the optic nerve, and were certainly due to the 
over-doses of atoxyl given. 

Thiroux ET d’Anfreville, ‘ La maladie du Sommeil au 
Senegal/ record, ‘ trois cas traites, guerison dans un cas.’ Rapport 
de Laveran, (Bull. Acad. Med. Seance de 26 fev. 1907). 

HOLLEBEKE, ‘ Traitement de la trypanosomiase par l’atoxyl. 
Notes cliniques et therapeutiques/ (Bull. acad. roy. de med. de 
Belgique, Vol. XXI, 1907), treated eight Europeans for trypano¬ 
somiasis. Gave daily injections of 0'2 gramme of Atoxyl. Never 
observed any eye-symptoms. After six months, he stopped the 
treatment and the same improvement continues. He considers his 
cases as cured. 

JAKIMOFF, ‘ Zur Atoxylbehandlung der experimentellen Dourine,’ 
(Deutsche, med. Wochenschrift. 1907. No. i(>), cured white rats 
infected with Dourine by Atoxyl. 

MARTIN, in a paper entitled ‘ Maladie du Sommeil. Cinq 
nouveaux cas de trypanosomiase chez les blancs. Essais de traite¬ 
ment/ (Ann. de Inst. Past. Tome XXI, Mars, 1907), states that Atoxyl 
has a very noticeable action on the trypanosomes, which disappear 
from the blood, as in malaria the parasites disappear under quinine 
treatment. Concerning whether or not a permanent cure has been 
effected, he states that it is impossible to say anything as the time 
was too short. 

COOK, at a meeting of the Society of Tropical Medicine and 
Hygiene, Oct. 26, 1907, discussed the question of sleeping sickness 
in Uganda, and stated that Atoxyl was by far the best drug for the 
cure of the disease that had yet been tried, and ‘ that it was greatly 
to the credit of English investigators that an Englishman, Thomas, 
of Liverpool, first applied it to the treatment of sleeping sickness/ 

UHLENHUTH, 1 Demonstration von mit Atoxyl behandelten 
Dourinekaninchen * (Deut. med. Wochenschrift, 1907, No. 30); 



UHLENHUTH, HUBENER and WoiTHE, 4 Experimentelle Unter- 
suchungen liber Dourine mit besonderer Berucksichtigung der 
Atoxyl behandlung ’ (Arb. a. d. Kaiserl. Gesundheitsamte, Bd. XXVII, 
h. 2, 1907); 

UHLENHUTH, Gross and Bickel, 4 Untersuchungen liber die 
Wirkung des Atoxyls auf Trypanosomen und Spirochaten ’ (Deutsche 
med. Wochenschrift, 1907, No. 4), give the results of treatment of 
experimental animals infected with T. equiferdum (Dourine) with 
Atoxyl (a) preventive, (b) curative. Preventive results were not very 
encouraging. They were able to drive the Dourine parasites out of 
rabbits, rats and mice, and to keep the animals alive. The authors 

do not state whether it is a definite cure, as the time of observation 
was too short. 

Ehrlich’s experiences with Atoxyl in experimental treatment are 
very favourable. He was able to prove that the different trypano¬ 
some strains become after a time resistant to the drugs, and he got 
the ‘festen’ strains; an Atoxyl resistant, a para-Fuchsin resistant 
strain, which did not even in sub-inoculation react to the drug. 

The Atoxyl resistance may partly explain the unfavourable results 
in some cases of sleeping sickness, as described by Kopke and 
Broden. 

To FOURNEAU (Journal Phar, et Chim., 6ieme serie, T. XXV, 
1 April, 1907) is due the credit of showing that, chemically, atoxyl 
is not a new preparation, having been synthetized as long ago as 
1863 by Bechamp in the early days of the synthesis of aniline colours, 
fuchsin being produced in abundance at the same time. Bechamp 
supposed that he had in hand an anilide of ortho-arsenic acid. 
Fourneau supports this view, and the body is described as an anilide, 
but this from the evidence given by Moore, Nierenstein and Todd, 
and by Ehrlich and Betheim, is probably an error. 

EHRLICH, in a lecture delivered before the Berliner medizinischen 
Gesellschaft, February 13th, 1907 (Berl. klin. Woch. No. 9-12, 1907), 
stated that Atoxyl was the sodium salt of p-amido-phenyl-arsenic 
acid, with four molecules of water of crystallization. The analyses of 
Moore, Nierenstein and Todd (Bio-Chemical Journal, Vol. II, 
Nos. 5-6, 1907, pp. 300-374) yield the formula (NH 2 ) (C 6 H 4 ) AsO, 
0 Na 0 H, 3 H 2 0 , and they had independently come to the conclusion 
that the arsenic radicle was united directly to the ring. Since then 



Ehrlich and Bertheim have published the details of their chemical 
work, showing that the arsenic radicle is in the para position to the 
NH a group. 

Moore, Nierenstein and Todd show that Atoxyl is not, as it was 
originally described, an anilide of metarsenious acid, but is an 
exceedingly stable chemical substance with the arsenical radicle 
directly attached to the benzene ring. It was shown that the aqueous 
solution is strongly dissociated electrolytically, giving in consequence 
an apparently low molecular weight by the freezing point method 
and possessing a high electrical conductivity. Except on standing 
in aqueous solution, it is a most stable compound, and neither aniline 
nor arsenic are easily detachable from its molecule by chemical 
means. 

Its toxic properties are neither those of arsenic nor of aniline even 
when pushed to excess, and its therapeutic action is rapid; from this 
and its high conductivity, showing high dissociation, the conclusion 
was drawn that its activity must be ascribed, not to free inorganic 
ions or to free aniline, but to a complex organic ion containing both 
the arsenical and aniline radicles. 

Since the introduction of Atoxyl for the treatment of trypano¬ 
somiasis a large number of observers have tested it, and all are now 
united in giving it the premier position as a trypanocide. 

The results of most observers upon the treatment of sleeping 
sickness by Atoxyl alone completely confirm the results stated above 
as having been obtained by Thomas and Breinl in the experimental 
trypanosomiasis of animals. Thus, cases have been described which 
were apparently permanently benefited and might be described as 
cured, but in a great many of them recurrences were observed, and 
finally the infection became persistent, the trypanosomes becoming 
‘ Atoxyl-fast * and being apparently no longer affected by the drug. 

Quite recently a second distinct advance has been made in the 
experimental therapeutics of trypanosomiases by workers of the 
Liverpool School. This consists in treating infected animals, from 
which the trypanosomes have primarily been driven out of the 
blood by the use of Atoxyl, by a second drug, so as to prevent the 
recurrences which so often follow Atoxyl treatment alone. 

The general principle underlying the combined method of treat¬ 
ment by two successive and quite different drugs is that when an 



infective organism, such as a protozoon, shows two distinct phases in 
its life-history, then these two phases ought to be attacked by 
separate drugs, and it is not only possible but probable that a drug 
which affects the first will not affect the second, and vice versd. 

The application of this bio-chemical principle, which has led to 
success in the prevention of recurrences in trypanosomiases, may be 
very wide in experimental therapeutics, especially in protozoon 
diseases where different phases in the life-history occur in nearly all 
cases. The two drugs necessary to attack two successive stages of 
the parasite will certainly not always be found to be an arsenical 
compound and a mercurial compound ; but it is established as a 
principle that given a parasite has two successive phases, A and B, 
then the problem of experimental therapeutics is to find two remedial 
agents, a and b , of which a kills phase A and b kills phase B, and 
then to apply the remedies a and b in succession, killing off phase A 
as completely as possible with remedy a , and then attacking phase B 
with remedy b t and continuing this rotation until the animal is free 
of infection. 

The research on the combined treatment was commenced in the 
Bio-chemical Laboratory of the University of Liverpool by B. Moore, 
Nierenstein and Todd in October, 1906, and a preliminary report 
published in March, 1907, followed by a fuller account in May, 1907; 
the work is now being continued on large animals in the Runcorn 
Laboratory, and, as far as cases are available, upon human trypano¬ 
somiasis. 

At the time of the commencement of this research, a second 
phase in the life cycle of the trypanosome was not known with 
certainty to exist, but since then the life-history has been more 
completely investigated by J. E. Salvin-Moore and Breinl, and these 
observers have clearly shown the existence of such a phase. 

The existence of this other phase was suspected by Thomas and 
Breinl, Moore, Nierenstein and Todd, from the regular way in which, 
after the trypanosomes had been completely driven out of the 
peripheral blood by Atoxyl, recurrence again took place, although 
the investigation of organs and tissues other than the blood had 
failed to demonstrate anywhere a storage of the ordinary phase of 
trypanosome. 

On these grounds, these observers determined to first drive out 



3i 


the parasite from the peripheral blood by Atoxyl, and then when the 
blood was free to treat by other drugs, paying no attention to whether 
these drugs had any effect upon the usual pffiase of the trypanosome 
or not. 

Accordingly a series of experiments was commenced, in which 
rats infected experimentally with Trypanosoma brucei were used on 
account of the rapidity of action of this type of trypanosome. After 
driving out the trypanosome with Atoxyl, the salts of the different 
heavy metals were given ; salts of Silver, Lead, Copper and Antimony 
respectively were employed without any marked results, but when 
Mercury was used in the form of the bichloride a distinct beneficial 
result was at once obtained. While the entire series of controlled rats 
treated with Atoxyl alone succumbed, nearly 70 per cent, of the rats 
given the double treatment survived, never showing any recurrence 
of trypanosomes, and of the remaining 30 per cent, only 8 per cent, 
showed recurrences of trypanosomes. 

It may be emphasized that the Mercury salt alone has not the 
slightest effect upon the ordinary phase of the trypanosome as seen 
in the peripheral circulation. This appears to demonstrate clearly 
that the two drugs act upon two quite distinct phases. 

Similar results are at present being obtained with other classes 
of animals, and indications of like results with the more slowly acting 
7 rypanosonia gambiense of sleeping sickness. , 

The two drugs are likewise being employed in the treatment of 
sleeping sickness in man, and the results so far obtained are distinctly 
encouraging. 

PLIMMER and THOMSON, ‘ A Preliminary Summary of the Results 
of the Experimental Treatment of Trypanosomiasis in Rats * (Proc. 

Roy. Soc., July 20th, 1907), got the best results by combining Atoxyl 
and the different Mercury preparations (Sozoiodol, Donovan's 
solution), and certainly some of the rats were cured. They also used 
Iodipine. Plimmer and Thomson were able to confirm Ehrlich’s 
experiments on atoxyl-resistance. 

Plimmer and Thomson, * Further Results of the Experimental Antimony 
Treatment of Trypanosomiasis in Rats’ (Proc. of the Royal Soc., 
read Nov. 7th, 1907), recommend for the treatment of sleeping 
sickness Potassium antimony 1 tartrate. Trypanosomes disappear 
very rapidly, but there is a bad effect on the rats treated. They 



su gg es t the use of Sodium antimonyl tartrate. Out of 25 rats which 
have been treated with this substance, 23 had no relapses from 25 to 
26 days. No local disturbances are caused by the injection of the 
drug into rats, in doses of up to 0*5 centigramme. 

In the hands of one of us (A. B.) Sodium antimonyl tartrate has 
not given the good results Plimmer and Thomson described. A 
fairly virulent strain of T. equiperdum was used. Out of twelve rats 
treated with two to four injections of 0*25 c.c. of a 1 per 
cent, solution (0*35 of the same solution was the fatal dose) 
only one rat which was treated during the incubation period 
of the disease is still alive, all the others having died ten to 
fifteen days after the last injection of the drug, their blood 
swarming with parasites. One horse infected with a strain 
of cattle trypanosomes, brought back from the Congo, has been 
under treatment since December 9th, having received five intra¬ 
muscular injections, ioc.c. of a 1 per cent, solution of Sodium 
antimonyl tartrate. It is still showing parasites in a very small 
number from time to time; they were seen even three days after 
the last injection. The local effects of the drug in rats are very 
severe : necroses and sloughing. 

The parasites, however, disappear very rapidly indeed, generally 
speaking more so than after a corresponding injection of Atoxyl. 

But notwithstanding our results, we consider that the introduction 
of another metal (Antimony) belonging to the same group as Arsenic 
is a further progressive step in the treatment of sleeping sickness, 
and is moreover very suggestive. 



MY EXPERIENCE OF TRYPANOSOMIASIS 
IN EUROPEANS AND ITS TREATMENT 
BY ATOXYL AND OTHER DRUGS 


BY 

Sir PATRICK MANSON, K.C.M.G., F.R.S., &c. 

{Received January 23rd , 1908) 

In view of the recently recorded experiences of Campenhout, 
Broden, Kopke, Koch and others in the treatment of human 
trypanosomiasis, especially in negroes, by atoxyl, it may not be 
inopportune if I gave some account of my experience of this disease 
in Europeans, and of atoxyl and other drugs in its treatment. 

My experience of trypanosomiasis in man extends to seventeen 
cases—seven negroes, ten whites. The negroes, who had been 
brought to Europe for purposes of clinical study, and because they 
had already entered on the terminal phase—sleeping sickness—of 
the infection, all died. They did not have the benefit of Thomas’s 
important discovery of the therapeutic value of atoxyl. I shall not 
allude to them further. 

Of the ten whites, three of the cases have been recorded already 
by myself and others. For the sake of completeness I shall briefly 
mention here these three cases, along with the seven unrecorded 
cases, giving them along with the latter in the order in which they 
came under my observation, but referring the reader to the medical 
journals for details. 

I.—Mrs. H. M. was first seen by me on July 17th, 1901. She was 
then 40 years of age, and had resided on the Congo for two periods 
of two years and one year respectively. During the latter period she 
had suffered much from fever. Being pregnant at the time she came 
home, arriving in England in April, 1901. She had fever all the way 
home. A week after her arrival her child was bom, and from that 
time till the date of her visit to me she had attacks of fever lasting 
for three days at a time and occurring at intervals of seven days 
with considerable regularity. She also suffered with pains in her 

D 



34 


hands, ankles and knees, for which she took sodium salicylate with 
some relief. She informed me that the attacks of fever were preceded 
and accompanied by a circinate erythematous eruption on her face, 
limbs and trunk, and that this tended to subside with the subsidence 
of the fever. She was anaemic and her skin showed traces of the 
erythema she referred to. The spleen and liver were not palpably 
enlarged. 

As the trypanosoma had not been discovered at that time I 
regarded the case as one of malaria, and prescribed a systematic 
course of quinine. 

I saw this lady again on April 9th, 1902. She told me she had 
improved, notwithstanding the fatigue consequent on the illness of 
her baby which had died on December 19th, 1901 (? trypanosomiasis). 
Three weeks before her visit to me she had caught a chill and the 
fever, which had been in abeyance for a long time, had returned. 
During this period of three weeks she had had three attacks. 
Nevertheless, as compared with her condition in July, 1901, she had 
put on flesh and no longer appeared anaemic. She informed me that 
the patches of erythema still showed themselves at times, but were 
less pronounced than formerly. 

I did not see this patient again at this time, but learned that she 
had an attack of irido-cyclitis and that subsequently she returned 
to the Congo. 

In the autumn of 1902, after having learned from Dutton’s case 
and from Case No. 3 (J:o be presently alluded to) that irregular fever 
along with erythema multiforme were in a patient from tropical 
Africa probably symptomatic of trypanosomiasis, I wrote to her 
husband on the Congo requesting that an examination of this lady’s 
blood be made for trypanosomes. Before receiving a reply to my 
letter I heard from Dr. Broden that he had examined her blood and 
had found the parasite. 

Dr. Broden put her on arsenic (Fowler’s solution). The case did 
well. Fever and trypanosomes disappeared. When I saw her during 
a subsequent visit to England in 1906-7, by way of encouraging them, 
I showed this lady to two trypanosomiasis patients I had at the time 
under treatment as an example of recovery from the disease. She 
was stout and healthy looking, and was free from all symptoms of 
the infection. I hear she is still in excellent health. 



35 


This patient must have received her infection some time during 
1900. The conclusions that she has overcome the infection and that 
trypanosomiasis in man is not necessarily fatal are, it seems to me, 
justifiable. 

Dr. Broden has published his notes of this case. 

II. —H. K. This was Forde’s original case in which Trypanosoma 
gambiense was first definitely recognised (and for the first time in 
human pathology) by Dutton. It has been fully described by Dutton 
in the publications of the Liverpool School of Tropical Medicine, 
and elsewhere by Forde. I mention it here as it constituted my first 
conscious experience of trypanosomiasis in man. I saw the case in 
August, 1902, and had the clinical points of the disease demonstrated 
to me by Dutton. It was from what I saw on that occasion that I 
was enabled to recognise clinically the disease in the next case. The 
medicinal treatment consisted principally in the administration of 
arsenic, quinine and urotropin. The patient died the following 
January, about one year and eight months after the presumed date 
of infection. 

III. — Mrs. S. was seen by me for the first time in October, 1902. 
She presented the usual clinical picture of trypanosomiasis, and the 
parasite was found in her blood. The case has already been fully 
recorded in the British Medical Journal of May 30th and December 
6th, 1903, and elsewhere. I may mention here that the first 
indication of the disease occurred in August, 1901, supervening, 
apparently, on an insect bite on the leg. The patient died of sleeping 
sickness on November 26th, 1903, two years and three months after 
infection. The treatment included arsenic, quinine, methylene blue, 
and many other drugs, but not atoxyL 

IV. — w. z., an engineer on one of the lake steamers in Uganda, 
came under my observation on October 9th, 1905. His story was 
that early in the year he broke his leg; that on this account and 
because he suffered from fever he had been in hospital in Uganda for 
a considerable time; and that trypanosomes having been found in 
his blood he was invalided on July 26th. On arrival he went to his 
home in Scotland, where, with the exception of two days’ fever, he 
kept well and put on flesh. He stated, however, that he had suffered 
from dull pains in his legs and that once he had a swelling in his 
left foot. 





When I saw him, there was a well-marked circinate erythema on 
his chest, and the cervical and inguinal lymphatic glands were 
enlarged. On examination of the blood trypanosomes were found. 
His pulse was rapid—108—his spleen palpable, knee jerks 
exaggerated, slight right ankle clonus. He complained that he felt 
weak, and also of subjective symptoms of numbness in the legs. 
Otherwise he appeared to be well. Blood count 4,000,000. He was 
sent to hospital. 

During his stay there he had a malarial attack and benign tertian 
parasites were found in his blood. Quinine quickly got rid of this 
infection. He also developed a specific periostitis which yielded to 
potassium iodide. 

From October 18th, 1905, to January 25th, 1906, he was treated 
by trypanroth and, later, by trypanroth and arsenious acid, the latter 
hypodermically. The erythema persisted, however, or, if it faded for 
a time, would again return. Occasionally the erythematous spots 
gave one the impression that they were slightly oedematous. 
Trypanosomes were also occasionally found in the blood, but gland 
puncture, which was twice practised, was negative. The temperature, 
except during the malarial attack, remained normal throughout. 

The skin and urine became deeply stained by the trypanroth. 
On January 26th, albumin having appeared in the urine, the 
trypanroth and arsenious acid were stopped. On February 5th 
treatment by atoxyl injections was commenced. Beginning with one 
grain twice a week it was gradually raised to four grains twice a 
week, at which dose it was continued till the spring of 1907. 

Very soon after commencing the atoxyl he became conscious of 
an improvement in his general health. The erythema no longer 
showed itself on the trunk, and for over six weeks, notwithstanding 
frequent and prolonged search, trypanosomes could not be found in 
his blood. He now insisted on leaving hospital, promising to continue 
treatment and to report from time to time. 

He returned to his home, where he kept in perfect health and 
gained weight. Wishing to get reinstated in his former appointment 
in Uganda, he came to London during the autumn of 1906. His 
blood was again carefully examined for trypanosomes, but n6ne were 
found. However, a monkey injected (25th August) with his blood 
developed trypanosomiasis and died. 



37 


Once more he returned to Scotland and continued the injections, 
his health keeping satisfactory in every respect. Early in 
1907 he again came up to London. Again his blood was 
carefully examined with negative result A monkey injected 
with the blood failed to develop trypanosomiasis. It was not 
considered prudent to allow him to return to Uganda, but he was 
encouraged to seek employment elsewhere, and when an appointment 
offered in the West Indies he left England in the early spring of 
1907. On the voyage out he was shipwrecked, and during eleven 
days suffered great hardships, wandering about the mangrove swamps 
at the mouth of the Magdalena river. Nevertheless he kept in 
perfect health, and when I saw him on his return on 19th April, 1907, 
he appeared healthy and robust, without trypanosomes in his blood 
and with no sign of trypanosomiasis about him. As he had lost his 
syringe and atoxyl when shipwrecked, he had had no injections for 
many weeks. 

Subsequently he again set out for the West Indies, and when last 
heard of was still in perfect health. 

V.—J. M., a botanist and agricultural expert, was in British 
Central Africa from June, 1897, to September, 1899. I examined 
and passed him for service in Uganda in March,. 1901. He was then 
30 years of age and in good health. 

He arrived in Uganda on June 17th, 1901. He informed me that 
before crossing the lake he had several small fevers (probably 
malarial) at Kusumu. He also informed me that a fortnight after 
his arrival in Etebbe he was bitten by * something * in front of his 
left ankle; the part swelled and he had fever about the same time. 
With this exception he kept well for a year. About June, 1902, he 
began to ail—languor, loss of appetite. At Christmas of that year, 
and on and off till March of 1903, he had several heavy fevers, 
accompanied by cerebral symptoms. He became melancholic, had 
delusions of persecution, and at one time was suicidal. Trypanosomes 
were found in his blood and he was invalided home. 

I saw him on August 1st, 1903. He had no fever then, and 
looked fairly healthy, but his manner was strange and there were 
definite trypanosoma erythema patches on his trunk and I found 
trypanosomes in his blood. 

I saw him subsequently from time to time. Occasionally he had 



puffy erythematous patches on his face and trunk, and once a big, 
swollen red and very tender patch on one gluteal region. 
Occasionally he had slight fever, and trypanosomes could usually be 
found in his blood. A principal complaint was of weakness and 
stiffness of the legs. 

He was subjected to a variety of treatments, including arsenic 
and trypanroth, but not, so far as I know, atoxyl. 

I lost sight of him. I heard that ultimately well marked sleeping 
sickness symptoms set in, and he died in University College Hospital 
in 1906. 

VI.—Mrs. R., aged 31, had resided on the Congo (Bogandango) 
from 1898 to 1902, and, with the exception of fever for three days, 
during all that time had enjoyed good health. After furlough in 
England she returned to the Congo in September, 1903. She kept 
fairly well till June 15th, 1905, when she had a severe attack of 
abdominal pain, vomiting and diarrhoea. These symptoms recurred 
three days later (June 19th), when they were accompanied by fever 
and, on the following day, by haemoglobinuria. The haemoglobinuria 
persisted for five days. During convalescence she noticed that her 
right ankle had become enormously swollen, purplish in colour and 
very painful; at the same time a gland in the corresponding groin 
enlarged to the size of a hen’s egg. Fever returned on June 26th 
and subsequently, off and on, about every four or five days, till 
September, when, being greatly debilitated, she was ordered home. 
On her way down river she was examined by Dr. Broden at Leopold¬ 
ville, who found trypanosomes in her blood, besides enlarged cervical 
glands and a rapid pulse (120) with a normal temperature. 

I saw Mrs. R. on October 9th, 1905. She was then very much 
emaciated, feeble and sallow. There were patches of ringed 
erythema on her chest and flanks and some enlarged cervical glands 
(right supra-clavicular). The spleen also was enlarged, and over the 
right ankle I found some reddish staining of skin, the remains of 
the inflammation in June. Trypanosomes were present in the blood ; 
blood count 3,700,000. 

She was sent to hospital and treated with arsenic and, later, 
Donovan’s solution. On November 22nd she commenced atoxyl 
injections up to two grains twice a week. Over that dose the drug 



A<) 


seemed to cause nausea. Relatively the dose was a medium one, as 
she weighed only ninety pounds when treatment commenced. 

On November ist her right ankle became swollen and so painful 
she could not walk; but from the time atoxyl was commenced 
improvement set in and persisted The erythema and the trypano¬ 
somes disappeared; temperature, which had been variable, became 
steady and normal, and her weight rapidly increased. Within a 
couple of months, from being sallow and emaciated she became ruddy 
and plump, and felt in excellent health. 

After her discharge she returned from time to time to hospital to 
show herself and to have her blood examined. Trypanosomes were 
not found again; the erythema did not return; monkeys injected 
(January 12th, 1906) with her blood were not infected (March 29th, 
1906), and good health continued. She left for the Congo on 8th 
March, 1907, against advice, but promising to continue the atoxyl 
injections. 

I am informed that a letter, dated nth September, 1907, had 
been received from her husband in which it is stated that ‘ she has 
been getting stronger and has been able to do quite a lot of language 
work. But she has to take great care, for evidently there is 
something causing a rise of temperature occasionally.’ Soon after 
her arrival on the Congo she had a haemorrhage of some sort which 
pulled her down very much; apparently, she has taken some time to 
recover from this; possibly, judging by the temperature, the 
trypanosomes are again active. 

VII.—H. C. C. S., aged 36, an engineer, arrived at Benguella, 
Portuguese West Africa, in June, 1904. His work took him up 
country some 100 to 150 miles from the coast. He had his first 
fever the following November. Attacks recurring very frequently, 
he had to be invalided, and arrived in England on May 29th, 1905. 
Soon after landing he had two attacks. I first saw him on June 23rd. 
He had no fever at the time, but he was anaemic and his spleen was 
enormously enlarged. I put him on quinine, 15 grains every tenth 
and eleventh day. Notwithstanding these doses fever kept recurring 
every few days. My locum tenens saw him on August 15th and 
ordered him five grains of quinine three times a day, apparently 
with benefit, for in a note dated August 24th it is stated that he had 
no fever, and that the spleen could no longer be felt. Subsequently 



the patient married and returned to Benguella during the autumn 
of 1905. 

For a time he kept fairly well and did much hard work in the 
interior on railway construction. He remembers that about this 
time he was bitten or stung on the leg by some unrecognised animal, 
supposed to be a centipede or scorpion. The part swelled and was 
very painful. 

Fever now returned and, in addition, he got dysentery. Between 
the two he became so ill that he had once more to return to England, 
where he arrived on February 17th, 1906. 

I saw him the same day. He was in bed. He had no fever at 
the time, but he was intensely anaemic, emaciated and weak, and 
evidently very ill. Both spleen and liver were much enlarged, the 
latter being tender as well. He was passing from four to five 
dysenteric stools daily. Quinine he said, aggravated the dysentery 
and gave him severe gastralgia. I examined his finger blood but 
found no malaria or other parasites in it. As he had not taken 
quinine recently, I was surprised at the absence of malaria parasites. 
I saw him daily, and on 22nd February, observing that the spleen 
had undergone a sudden increase in size, and from this suspecting 
an impending malarial attack, I gave orders that I should be sent 
for so soon as temperature rose. A few hours later I was summoned. 
The temperature was then 103°. I took blood films and found in 
them, not malaria parasites, as I expected, but considerable numbers 
of trypanosomes. I now made a careful examination of the skin 
and lymphatics, and recognised several characteristic patches of 
erythema and at least one definitely enlarged posterior cervical 
gland. Temperature rose on this occasion to 106° but quickly fell, 
although for three or four days some fever of a remittent type 
persisted. Trypanosomes were found for some days, their numbers 
gradually becoming fewer under the atoxyl treatment which was at 
once instituted. 

From time to time, at intervals of a few weeks, although the 
general condition of the patient, including the dysentery, steadily 
improved, there were short recurrences of fever, each recurrence being 
associated either with the appearance of trypanosomes, or of benign 
tertian malaria parasites in the blood ; so that without the microscope 
it was impossible to say whether a given relapse was trypanosomal 



or malarial in nature. On the discovery of the malarial infection 
quinine was given systematically at definite intervals as well as 
atoxyl, and was now very well borne. 

At first the atoxyl was given in one grain doses (io per cent, 
solution) hypodermically every third day. It was gradually raised to 
2*3 grains, beyond which, after not a few attempts, it was impossible 
to push it. Every time a 2*5 grain dose was given, violent and 
alarming gastralgia ensued. The 2*3 grain doses of atoxyl, and 
occasional 10 grain doses of quinine were therefore steadily 
persisted with. 

Under the persevering use of these drugs and with careful 
nursing the patient slowly improved, the erythema, the adenitis and 
the dysentery disappearing. When the weather became milder, and 
he could walk about, the patient was removed to a healthy and 
bracing place in the country, where I saw him from time to time. 
There the febrile attacks became milder, returning at longer intervals, 
the trypanosomes being found only occasionally in the blood and 
never in large numbers. In the course of the summer of 1906 he 
suffered at one time from severe dental neuralgia, and twice from 
smart attacks of orchitis. Notwithstanding this, general improve¬ 
ment continued. He spent the winter of 1906-7 on the high Alps, 
where, with the exception of a brief but painful attack of what 
might have been erythema nodosum in one leg, he kept quite well 
and gained strength. I saw him again on July 25th, 1907, and noted 
that ‘he had had no fever since 21st February with the exception 
of that attending a slight cold in May (temperature ioi°). Weight 
9^ stone—the highest he has ever been in his life. Feels quite well/ 

He refused to consider himself any longer an invalid. A 
situation was offered to him in South America, my consent being a 
condition of the appointment. This I promised provided injection 
of his blood into monkey, rat and guinea-pig proved negative. These 
injections were made in August. All the animals were alive and free 
from trypanosomes in October when he sailed for South America in 
the best of health, promising to keep up the atoxyl injections for 
another year and to report progress. 

A letter just received and dated 31st December, 1907, stated that 
‘he is very well indeed/ and ‘to see him now no one would think 
he had ever had an illness.’ 



4 - 

VIII. — C. G., aged 34, an engineer, was stationed at Katanga 
on the Upper Congo (Lualaba river) for three and a half years. He 
took five grains of quinine daily and enjoyed excellent health till 
November, 1906. About that date he began to suffer from fever, 
apparently uncontrolled by quinine. On November 9th, his 
temperature at the time being 102°, he started to return to England 
via Rhodesia and the Cape, riding 500 miles of the way on a bicycle 
and suffering from fever all the way. On the voyage from the Cape 
he gave up taking quinine as it seemed to do him no good. Fever 
persisting, he became bilious and yellow. 

I saw him on his arrival (January 5th, 1907). His temperature 
was 103°, pulse 112. He was somewhat emaciated, slightly icteric 
and markedly anaemic. Liver and spleen, especially the latter, were 
both enlarged. The superficial cervical glands were also slightly 
enlarged and the skin of the trunk was splashed with rings and 
patches of erythema multiforme. A blood examination gave 
2,848,000 erythrocytes per c.mm. and 4,000 leucocytes per cmm., the 
large mononuclears being in marked excess (37*3 per cent). Malaria 
parasites were not found at that time, but trypanosomes were 
numerous. 

A/oxyl .—He was at once put to bed and atoxyl in hypodermic 
injections begun. At first the injections were given every second 
day, the dose being rapidly increased from 1} to 7 grains, and 
occasionally 8 grains. In a few days trypanosomes had disappeared 
from the peripheral circulation, so that when Dr. Todd, of Liverpool, 
saw the patient with me on January nth he failed to find a single 
specimen during a prolonged examination. The skin eruption, the 
adenitis, the fever and, in great measure, the debility disappeared 
equally rapidly. On January 13th and 14th there was a return of 
fever which, on microscopical examination, proved to be malarial 
benign tertian. Quinine was now given in addition to the atoxyl 
and repeated at intervals ever since. 

Except during four short periods the atoxyl injections have been 
continued. They were intermitted from May 26th to June 12th, from 
June 26th to July 9th, and from July 16th to July 25th, when he was 
taking perchloride of mercury; and also from November 6th to 
November 27th (three weeks), when he was being treated with 
sodio-tartrate of antimony. The dose of atoxyl varied from 2 \ to 



43 

9 grains ; latterly it has been 3 grains every second day. At no time 
has there been any local reaction or sign of arsenical poisoning. 

Perchloride of mercury .—Notwithstanding several febrile attacks 
and frequent and careful examination of the blood, no trypanosomes 
appeared in the peripheral blood till May 21st. On that day a few 
were found. Atoxyl in 8-grain doses was being administered at the 
time. The same dose was repeated on the 22nd, 24th, 26th and 
28th. The trypanosomes having disappeared, hypodermics of 
perchloride of mercury in | per cent, solution were commenced and 
repeated daily for fourteen days, the dose being rapidly increased 
from 15 to 30 minims (about one-seventh of a grain), at which it was 
kept for five days, when atoxyl was resumed in 2^-grain doses every 
second day. * A week later there was another febrile attack 
concurrent with a fresh invasion of trypanosomes. The dose of 
atoxyl was now increased to five grains. After four injections of 
this strength, and the trypanosomes having disappeared, the 
perchloride was resumed from June 27th to July 6th. On July 10th 
there was a rise of temperature to ior6°, but trypanosomes were not 
found, though carefully searched for; the atoxyl was again resumed. 
The perchloride injections having caused much pain and irritation, 
Hydrarg. c creta, one grain three times a day, was substituted and 
continued from the 15th to the 22nd of July, when, the gums being 
slightly tender, it was stopped and the atoxyl resumed. On August 
nth temperature rose to 1017°, and trypanosomes once more 
appeared in the blood. 

Parafuchsin .—Treatment with parafuchsin (kindly suggested and 
supplied by Professor Ehrlich) was begun on August 16th, atoxyl in 
2 2> an d occasionally in 8-grain doses every second day being 
continued at the same time. Beginning with 5 grains, the dose of 
parafuchsin was gradually raised by 5 grains at a time till 20 grains 
were taken three times a day by the mouth in cachet. These large 
doses, though continued till October 13th (nearly two months), caused 
no disturbance. There was no intestinal irritation, nor, although the 
urine and sweat were coloured by the drug, was there albuminuria or 
urinary irritation. Trypanosomes, which were present when the 
parafuchsin was commenced, persisted till August 29th—a fortnight. 
After this, though looked for almost daily, they disappeared for a 
time. 



44 


The last dose of parafuchsin was taken on October 17th. Ten 
days later there was a rise of temperature, and once more the 
parasites appeared. About this time the patient apparently got a 
chill while salmon fishing and there was some rise of temperature, 
which was repeated on November 1st and 2nd, when a very large 
influx of trypanosomes was noted 

Sodio-tartrate of antimony .—On November 8th I administered a 
hypodermic injection of sodio-tartrate of antimony (kindly supplied 
by Mr. Plimmer)- i a grain, and followed it up with injections of 1, 
i£, 2, and 2 grains on the 10th, nth, 12th and 15th respectively. 
There was no nausea, intestinal disturbance, or albuminuria following 
the injections, but the local pain and irritation were great The 
parts became puffy, then hard, then fluctuating. One swelling, which 
had become very tense and discoloured, and seemed as if about to 
rupture, I incised About an ounce of dark sanious fluid escaped, 
and after a week an ash-grey slough was seen at the bottom of the 
wound. The slough took a fortnight to come away, and the wound, 
which never suppurated, took a very long time to heal. The other 
swellings took several weeks to subside. The pain consequent on 
the injections was very great, demanding morphia. It would subside 
towards morning and be comparatively in abeyance during the 
forenoon, but each afternoon it would wake up again, being most 
severe during the night, rendering sleep impossible. There was very 
little rise of temperature. The pain consequent on an injection 
lasted about a fortnight. 

It was remarked both in this and in another case that although 
given in different strengths to 1 per cent.) the weaker solution of 
antimony was quite as painful as the stronger; and, also, that 
intramuscular injection, though equally painful at first, caused less 
swelling than subcutaneous injection. 

When it became evident that hypodermic injection of antimony 
was impracticable, the drug was given by mouth to the extent of 
two grains a day diluted in two or three pints of water. This, along 
with three grain doses of atoxyl, is being continued. It causes no 
nausea or irritation of any sort. 

Up to the date of this note there has been no return of fever. 
The patient is leading an active country life and is feeling perfectly 
well. 



45 


IX.—W. R. E., aged 46, was in the West Indies from 1881 to 
1903, exploring and planting. 

In 1903 he went to Northern Nigeria as a Forestry Officer. 
Although his duties took him all over the country and exposed him 
to much hardship, he enjoyed excellent health during his first tour 
of twenty months. After furlough in England he returned to 
Northern Nigeria in October, 1905, and kept perfectly well till 24th 
September, 1906. About the latter date he got an irregular fever 
which resisted quinine in full doses. Not improving he was invalided, 
and arrived in England on 29th November, 1906. He had slight 
fever on the voyage home, and at that time observed certain red 
patches on his skin, which he attributed to pressure from his clothes. 
After six weeks in the country, during which he took long walks and, 
with the exception of a slight fever at Christmas time, felt very well, 
he was sent to me officially for report. 

I saw him first on January 15th, 1907. On examination I found 
his trunk covered with rings and patches of erythema; several 
cervical, axillary and inguinal glands were enlarged though painless; 
spleen and liver were also slightly enlarged and his pulse was quick 
100. I found trypanosomes in his blood. There were no other 
symptoms of trypanosomiasis or of other disease. He said he felt 
fairly well. I sent him to hospital for treatment and observation. 

Atoxyl injections were begun on January 25th, 1907. They 
were given every second day, the dose being gradually raised to 
seven grains. He remained in hospital from January 21st till March 
4th, 1907. For the first fortnight temperature was somewhat erratic, 
ranging from 98° to 99 0 F.; once it was ioo*8°. Subsequently, with 
the exception of one short rise of a few hours to ior8° on March 
5th, and attributed to a cold, temperature was steadily normal or 
subnormal. Erythema and trypanosomes disappeared about 
February 12th. 

After leaving hospital he returned to his home in the country, 
where the injections were continued. About July symptoms of 
peripheral neuritis—not of a very pronounced character—showed 
themselves. He was again taken into hospital and a course of 
mercury substituted for the atoxyl. The gums were slightly 
1 touched.’ The neuritis subsiding, the atoxyl was resumed in 
smaller doses. He now became so well that he wished to return to 



Africa. The atoxyl was stopped for a fortnight with a view to 
testing his supposed cure by injection of his blood into animals. At 
the end of the fortnight trypanosomes were again found in the blood 
and the atoxyl resumed. I saw this patient lately (16th January, 
1908). He says he never felt better in his life; he looks quite well, 
and no indication of trypanosomiasis can be detected on careful 
examination. 

X.—A. P. P., an engineer, aged 35, arrived at Boma, Congo River, 
on June 22nd, 1907. He was then in perfect health, and proceeded 
at once up country to a point about 150 miles above Stanley Pool, 
where he and his companions camped on the river bank from June 
28th till July 17th. Four days after his arrival there he began to 
ail with anorexia, depression, languor, drowsiness, and on 12th July 
took to bed with fever (102°). On July 17th his temperature had 
reached 107°. Next day he was brought to Leopoldville (104°), and 
on the following day—19th July- -Dr. Broden found trypanosomes in 
his blood. He received a large dose (I understood 1*5 gramme) of 
atoxyl hypodermically on the 20th. This gave rise to violent 
gastralgia, but on the 21st temperature had become normal and has 
remained so ever since, with the exception of slight brief rises (99*5 
to 102*4°) which recur with some regularity every fortnight or three 
weeks. On 23rd July daily injections of *25 gramme of atoxyl were 
commenced and, with occasional intermissions of a day or two and 
increases of the dose to *5 gramme, were continued till his arrival in 
Liverpool about the middle of October. 

Trypanosomes were unusually persistent in the peripheral blood 
in this case. Dr. Broden found them when the patient was at 
Leopoldville every time he looked for them. Dr. Breinl, who 
examined him on his arrival in Liverpool, found them, notwith¬ 
standing intensive doses of atoxyl, on each of five successive days; 
and I found them whenever I examined the blood—that is, on every 
alternate day—between October 23rd and November 8th. 

When I saw the patient for the first time on October 23rd he 
informed me that he had had no cutaneous eruption, and that only 
once—during the initial fever—when one slightly enlarged gland was 
detected on the right side of the neck—any adenitis. An inflamed 
patch of skin on the dorsum of the right foot, which began on the 
20th of September, and which he attributed to prickly heat, had 



47 


almost disappeared when I saw him. The spleen was slightly 
enlarged, and he looked more anaemic than the blood count 
(4,000,000) indicated. Appetite was poor, and he felt weak and 
depressed. There was no palpitation or breathlessness, and no 
headache even when he had fever. The only pain he remembered 
was intense aching in the legs coming on every night and keeping 
him from sleeping; this disappeared when he left Leopoldville and 
has not recurred. 

On November the 8th, trypanosomes being present in small 
numbers in his blood and temperature being normal, for the first 
time during his illness the characteristic erythema showed itself on 
the skin of the trunk. At the same time and for a day or two 
previously he had been profoundly melancholic, so much so that I 
feared the nervous system was becoming implicated and that the 
terminal phase of the infection was about to set in. He was so 
depressed I was afraid to allow him to go out alone. 

On the day the erythema appeared (8th November) I gave him a 
hypodermic injection of half a grain of sodio-tartrate of antimony, 
and also on the following days 1, i£, 2, and 2 grains respectively, and 
again after two days another 2 grains. The apparent effect of these 
injections was remarkable. By the third day the erythema had 
disappeared, his spirits had become good, and for the first time 
trypanosomes could not be found in the blood. This hopeful 
condition persisted till November 26th, when depression, though not 
so intense as on the former occasion, returned and trypanosomes in 
greater number than I recollect to have seen them in human blood 
were once more found. On November 26th and 27th he had two 
grains of antimony by the mouth. I was afraid to resume the 
antimony hypodermics on account of the intense irritation and pain 
they gave rise to. Given by the mouth the drug caused nausea and 
seemed to increase the depression. It was stopped, therefore, and 
atoxyl resumed. By the 30th trypanosomes had again disappeared 
and the patient was feeling much better. He left for New York on 
December 12th, with instructions to continue the atoxyl. A letter 
just received states that he is feeling much better. 

The more important facts (especially as bearing on treatment) of 
these ten cases are summarised in the following table: — 



4 « 


)ase. 

Where acquired. 

Duration. 

Treatment. 

Present 

state. 

1 

Congo . 

7 vears . 

Arsenic {Liquor arsenicalis) 

Well 

2 

Gambia. 

1 year 8 months 

Aisenic, quinine, urotropine 

Dead 

3 

Congo . 

2 years 3 months 

Arsenic, quinine, methylene 

Dead 




blue, etc. 


4 

Uganda. 

3 years . 

Arsenic, trypanroth, atoxyl 

Apparently 

well 

5 

Uganda. 

4 years . 

Trypanroth-arsenic. 

Dead 

J ! 


T J * " 



6 

Congo . 

1 

2 years 6 months 

Atoxyl . 

Apparently 


Benguela . 

2 years 3 months 

Atoxyl . 

well 

Apparently 




well 

1 

8 i 

Lualaba River 

1 year 3 months 

Atoxyl, parafuchsin, per- 

Apparently 


(Congo) 


chloride of mercury, anti¬ 

well 




mony 


9 1 

; 

Northern Nigeria 

1 1 year 4 months 

Atoxyl, perchloride of 

Apparently 




mercury 

well 

10 

Congo . 

6 months . 

Atoxyl, antimony . 

Improved 






Any conclusions we may be tempted to draw from these and 
other recorded cases as to the effect of treatment in human trypano¬ 
somiasis must be tempered by the consideration that we do not as 
yet know the limit of the duration of the infection in man, that it 
certainly may run a course of three or four years, and that the lower 
animals, especially the smaller laboratory kinds, as regards their 
reaction to drugs, form no very reliable guide to the action of the 
same drugs in man. 

Subject to these considerations we may provisionally infer that: 

1. Trypanosomiasis in man is not necessarily a fatal disease. 

2. Atoxyl has a marked effect in checking the clinical 
manifestations of the infection and in causing the parasites to 
disappear from the peripheral circulation. 

3. Notwithstanding continuation of atoxyl treatment, 
parasites may reappear again and again at uncertain intervals, 
and usually concurrently with a rise in temperature. 

4. Nevertheless, if the drug be persevered with, the parasites 
ultimately disappear for good and do not return. 





















49 


5* Large doses of atoxyl are not necessary to secure this 
result 

6. Large doses of atoxyl should be avoided, as they are apt 
to cause serious lesions, peripheral neuritis, optic atrophy, 
gastro-intestinal inflammation, and other toxic conditions which 
necessitate suspension of a valuable remedy. 

7. Trypanroth, mercury and parafuchsin seem ineffective in 
human trypanosomiasis. 

8. Antimony may have a therapeutic influence in trypano¬ 
somiasis, but the hypodermic injection of the sodio-tartrate is 
impracticable. 

The prospects of atoxyl treatment I consider most hopeful. As 
regards efficiency, promptness and mode of action, it seems to me 
that it is almost on a par with mercury in syphilis and quinine in 
malaria; and I think in using atoxyl we should conform our practice 
to what experience has taught us to be the best methods of using 
these other efficient and long tried remedies. 

I do not believe we can kill the trypanosome outright by one or 
two large doses of atoxyl, any more than we can kill the treponema 
of syphilis or the parasites of malaria by large doses of their respective 
specifics. Mercury does not immediately cure syphilis, nor does 
quinine immediately cure malaria; but they deprive the respective 
parasites of their pathogenic qualities and keep the patient alive and 
in good health till, in process of time, the parasites either die out or 
become permanently inert. So I read the action of atoxyl in 
trypanosomiasis, and so I would regulate its administration, being 
careful, as we would with mercury or quinine, not to push the drug 
too far, and thereby necessitate its suspension. Case 7 distinctly 
shows that a dose of 2*3 grains given twice a week controlled the 
disease; why then risk poisoning by a larger dose ? Some of my 
patients have been for months on 2 to 4 grain doses two or three 
times a week and have done well. 

I would therefore suggest for the routine treatment of trypano¬ 
somiasis, a two to three grain dose of atoxyl every second or third 
day and kept up for at least two years. At the same time concurrent 
specific disease such as malaria, syphilis, &c., should be carefully 
treated; and, further, the patient should be brought home to his 
native country, be spared fatigue, worry, exposure, excesses of all 



kinds and be placed in the most favourable hygienic conditions 
possible. 

I fear to overstate my opinion of the value of atoxyl given in 
this way in the treatment of trypanosomiasis. I am fully conscious 
that the evidence in its favour, though strong, is far from complete. 
However, besides direct evidence derived from its actual use in the 
disease itself, there is a good deal of collateral and indirect evidence 
derivable from its employment in other protozoal diseases—syphilis 
for example. Recently I had an experience of this kind which has 
greatly impressed me. The case was one of Kala-azar—admittedly 
an almost invariably fatal disease. The patient had been ill for 
many months. He had the usual hectic type of fever, was miserably 
emaciated, and had enormously enlarged spleen and liver; the spleen 
extended beyond the umbilicus. Liver puncture yielded the 
Leishman body in profusion, so that there could be no question as to 
diagnosis. He was given atoxyl injections over a long period, at 
first apparently with little benefit. Severe inflammation of gums, 
cheeks and palate, together with symptoms of peripheral neuritis 
setting in, the injections were stopped. Shortly afterwards the 
patient left hospital, in my opinion then, apparently to die. Soon 
after leaving hospital symptoms began to subside; he lost fever and 
sweats; he gained strength and appetite ; his spleen and liver shrank, 
and, when I saw him a week ago, he had been free from fever for 
five months and appeared to be quite well, although still a little 
weak. The liver was almost normal in size, and the spleen could 
just be felt under the ribs. Was this atoxyl? If so, there is hope 
now for the victims of yet another formerly hopeless disease. 

Before concluding this paper, there are one or two points I would 
like to draw attention to. 

It is not a little remarkable that of the ten cases of 
trypanosomiasis in Europeans which have come under my personal 
observation three of them were females. Considering the very small 
number of European females and relatively large number of 
European males in tropical Africa, this large number of females 
attacked with trypanosomiasis is a striking circumstance. I am 
dealing, it is true, with very small numbers, and it is quite possible 
that the relative disproportion I remark on is accidental; but when 
we reflect that whereas women in Africa expose themselves, as 



5 1 


compared to men, comparatively little to the conditions favouring 
the attack of Glossina pal pal is, the disproportion becomes still more 
striking. 

Another point that has attracted my attention in connection with 
these cases is the frequency (four in the eight cases in which the 
point was inquired into) with which the symptoms were immediately 
ante-dated by what was described as a bite on the leg. The biting 
animal may have been a Glossina, but in the case of females—and two 
of the bitten ones were females—one would suppose that the 
petticoat would afford a protection even more effective than the 
trouser does in men. 

Too much weight must not be attached to what may have been 
mere coincidence; but these facts are curious, and suggest further 
inquiry as to the possibility of some blood sucker, perhaps some 
species of house vermin, being an occasional vector of Trypanosoma 
gambiense. 




53 


REPORTS OF THE ‘SLEEPING SICKNESS’ 
EXPEDITION TO THE ZAMBESI 
FOR THE YEARS 1907-1908 

BY 

ALLAN KINGHORN, M.B., Toronto, 

JOHNSTON COLONIAL FELLOW, UNIVERSITY OF LIVERPOOL 
AND 

R. EUSTACE MONTGOMERY, M.R.C.V.S. 

OBJECTS OF THE EXPEDITION 

The objects of the Expedition were to determine to what extent 
sleeping sickness had invaded British South African Territory, to 
map out the distribution of tsetse flies and to advise on means for 
checking the spread of sleeping sickness. 

The Expedition was put under the charge of Dr. Kinghom and 
Mr. Montgomery—the latter paying special attention to Trypano¬ 
somiasis in animals. 

The first letter from the Expedition is dated November 22, 1907, 
from N’dola, North-Western Rhodesia, and is from Mr. Montgomery, 
relating to a correspondence between the members of the Expedition 
and Mr. Moffat, the local representative of the various Copper 
Properties in North-West Rhodesia and the General Superintendent 
of the Bechuanaland Coaling Co. 

Mr. Moffat, who has taken a very keen interest in the sleeping 
sickness question, has helped the members of the Expedition in 
every way. 

In a letter from Broken Hill, Rhodesia, dated October 23, 
1907, and directed to Mr. Montgomery, Mr. Moffat states the 
following in regard to the question of the tsetse fly and big game : — 
‘ I am especially keen on the question as to the connection between 
the game and the fly, though I am confident that there is a close 
connection, and that the fly will not exist long without the game, 
there are undoubtedly here and there spots where there is little or 
no game where the fly still remains. This possibly is only temporary; 
the fly will probably disappear from such places shortly/ Mr. Moffat 
then proceeds to discuss the question of establishing an Observation 
Camp at Broken Hill, asking for particulars of the probable cost of 
maintenance. In a letter dated November 21, 1907, from Broken 
Hill, Mr. Montgomery replied .- — 

E 



54 


‘ Broken Hill, Rhodesia, 

‘21 st November , 1907. 

1 Dear Mr. Moffat, 

* Your letter of 23rd ultimo reached me at N’dola. I am 
extremely glad that you realise the imperative needs of research into 
the “ fly ” question, for, as you will see, the success of this territory, 
commercial and industrial, will be extensively modified should good 
come of any preventative or curative measures. 

‘ I attach herewith a short draft of what I should consider the 
more salient points in the projected investigation and an estimate 
of the probable expense. But I would ask you to clearly understand 
that this estimate might, after a period of work, prove quite erroneous, 
for, as has been noted in this district already, there are several forms 
of “ fly ” disease, and the greater the number of varieties discovered 
the larger the expense of experimental animals, as each would demand 
a separate series in the treatment and morphological researches. 

‘ With regard to your kind suggestion that I should undertake 
this work should it ever receive sanction, I may say that it would be 
a real pleasure if it may be assumed that the question be approached 
in an earnest and scientific spirit, and that its inception would imply 
a free hand to do what was considered necessary to obtain results 
which would be, not only of scientific and indirect, but also of 
practical and direct value; but I do not wish to associate myself with 
any programme which would demand the waste of several months 
and not include in its object scientific research. 

‘ I am positive that for such an undertaking the Liverpool School 
of Tropical Medicine would lend their whole support, and though I 
do not anticipate any movement on your side until the completion of 
the tour upon which I am at present engaged on their behalf, yet 
should arrangements be made more rapidly no difficulty would arise, 
for the results to be striven for are in both cases identical. 

‘ Yours truly, 

‘R. C. Montgomery.’ 

‘ An investigation into “ fly ” disease, having for its object a 
curative or preventative remedy, would embrace the following main 
points, each of which could be approached more or less independently, 
whilst the results of each would overlap and serve as controls to those 
arrived at from other sources: — 



‘ I. The nature of the various varieties of “ fly ” disease parasites 
met with in domestic or wild animals in the territory; their effects 
upon domestic stock; and the action of certain selected drugs upon 
them, more particularly the employment of atoxyl and mercury and 
the use of the more easily obtained aniline dyes. 

1 2. The incidence of trypanosome infection in wild game; the 
nature and effect upon stock of the parasites found. 

4 3. An enquiry into the transmission of fly disease by means of 
biting flies, more particularly Tsetse, " Hippo ” and “ Blind ” flies, 
and the “ Stinging ” house fly* 

4 In the event of success attending the experiments at least one 
year would be required for thoroughly testing and proving the method 
of treatment; and since these would follow on those already 
conducted near Broken Hill, we commence from a working base 
which eliminates a considerable number of tentative experiments. 

‘ The other main points would receive attention concurrently. 

1 It is not to be expected that this period of time is sufficient to 
do more than elucidate a few of the many questions associated with 
biting flies and big game. One could only work from hand to 
mouth, and from deductions drawn, institute control experiments 
which might answer the questions—can flies other than tsetse 
originate an outbreak of disease? and can flies other than tsetse 
continue the transmission in a herd when once originated? 

‘ It is known that game harbour the parasite : we could only 
ascertain figures showing the proportion of each variety affected and 
the effects of these germs on domestic stock. To ascertain these 
with accuracy a very large number of head would require to be 
examined; but at any period of the investigation figures having 
certain values would be available. 

‘ I would then suggest one clear year as a minimum and estimate 
accordingly, leaving it to be understood that in research work it is 
impossible to precisely specify periods of time. 

1 The requirements of such an investigation would be : — 

‘ A main camp situated clear of all danger from contamination 
by wandering tsetse flies, in the vicinity of a plentiful water supply 
and grazing for experimental animals, preferably within easy reach of 


Hippo = Tabartus ; Blind = Haematapota; Stinging housefly = Stnmoxys. 



5 ^ 

transport facilities and allowing of suitable accommodation for all 
engaged. 

1 Could such a site be found, it would be desirable to place the 
main camp at a spot from which tsetse fly could be obtained in 
sufficient numbers to permit of transmission experiments being 
conducted there without the necessity of erecting a second station 
within a fly zone and rendering that fly proof; but for treatment 
experiments it is a sine qud non that no possible chance of re-infection, 
natural or accidental, should occur. 

4 The accommodation required would consist of (i) a laboratory 
building with doors and windows fitted, (2) an office room, (3) huts 
for the man in charge, (4) sheds for the use of both healthy and 
infected animals (certain of them to be rendered fly proof by means 
of wire mesh), (5) accommodation for all servants, (6) stores, buildings 
for small animals, netted runs for the same, &c., &c. 

4 All these could be made of timber and daub by native labour, 
and should not cost more than £150, excluding the imported doors 
and windows, the necessary gauze and wire netting, and possibly, 
sundry sheets of iron roofing in the event of thatch proving 
unsatisfactory, over either office or laboratory during the rainy 
season. The fittings of these buildings could be readily made from 
rough sawn timber, but should include at least one good case for 
books, papers and sensitive instruments. 

4 The laboratory equipment would be simple, and for ordinary 
purposes need not demand an initial expenditure of more than £100. 
Should it be found necessary to go more intimately into the 
morphology of either parasite or fly this amount might be doubled. 

4 The number of experimental animals would, of necessity, be 
uncertain; but, at the present moment, I consider that 50 head of 
cattle, 15 head of sheep and the same number of goats, 6 horses or 
donkeys, and several dozen dogs, monkeys, rabbits, guinea-pigs and 
rats would be sufficient. These would have to be obtained from a 
source precluding any chance of natural infection, and all precautions 
would have to be taken en route: factors which would slightly raise 
the cost. 

4 A vote of £500 would probably cover the purchases of all 
experimental animals. 

4 In addition, the sum of £50 should be added for the cost of 



57 


incidentals and sundries in the shape of buckets, gear, ropes, 
stationery, petty furnishing, &c. 

1 For the control and the maintenance of the animals and the 
upkeep of the station, a capitao, 2 cattle boys and say 20 ordinary 
men would be needed involving a monthly expenditure of about £20. 

‘This indicates that the sum of £1,000 is necessary, of which 
50 per cent, would be sacrificed in animals, and 25 per cent, for 
current expenses, but it must most clearly be understood that for the 
thorough investigation of such a wide field as the present subject 
embraces, as generous a treatment as is consistent with common 
sense should be given, and that a feeling of parsimony would tend to 
curtail necessary experimentation and seriously impair the obtaining 
of positive evidence which must be founded on proof derived from 
observation and experiments. It may be pointed out also that 
should no satisfactory results be obtained within the specified twelve 
months, only current expenses would continue until the estimated 
number of animals is exhausted, a circumstance which should not 
occur until the present views on the treatment of fly disease have 
been subjected to thorough test. 

4 For ascertaining the incidence of the fly parasite in wild game, 
I would suggest the establishment of sub-stations, or temporary 
camps, in say six districts of this territory; two at least of which 
should be in localities absolutely free from tsetse fly. At times 
convenient to the investigator, trips would be arranged to these 
localities and as many head of game as possible belonging to different 
species subjected to examination and inoculation into small animals 
which would be brought to the main camp for thorough observation. 

4 Should the main camp not be situated conveniently in regard to 
a plentiful supply of tsetse, a second, but more temporary, set of 
buildings would be required in a suitable locality. This would 
comprise fly-proof sheds in which to maintain both experimental 
animals and fly, and would involve the purchase of additional 
quantities of small mesh netting. 

1 To conclude, I would consider that a grant of £1,000 or £1,200 
would enable the main points known concerning fly disease in other 
parts of the world to be controlled here, and to test thoroughly the 
present suggested methods of treatment. Should it be necessary to 
enter the field of research more deeply this sum might prove 



5 * 


inadequate, and in any case, as already remarked, the investigator 
should be given as free a hand as possible and receive full confidence 
in the undertaking, and in no case should he be restricted in the event 
of the unforeseen occurring. To this sum would have to be added 
the expenses of the investigator and at least one intelligent assistant 
who, of necessity, need possess no qualifications.* 

In the following Memorandum Mr. Montgomery furnishes Mr. 
Moffat with a description of the extent and nature of Trypanosomes 
in cattle in the Broken Hill district. 

4 (i) Trypanosomiasis of cattle (fly disease) is very prevalent in 
the vicinity of Broken Hill at the present time. On all sides 
information relating to losses caused is accumulating, and the disease 
has been verified in 36 cases. 

4 (2) It is possible to isolate two distinct species of Trypanosoma 
from these cases: the clinical effects produced upon naturally affected 
cattle being similar, but the reaction in inoculated animals, particularly 
goats and sheep, differ. Both forms are distinct from the classical 
cases of T. brucei infection found in fly disease in Zululand. It is 
therefore possible that further research would show a greater number of 
trypanosomes pathogenetic to domestic stock and that the 
discrepancies noted and described by hunters and travellers when 
referring to the effects of the tsetse fly are due to this reason. 

4 (3) One of these trypanosomes, closely allied to T. dimorphon 
found in Senegambia, is fatal to cattle, sheep, goats, dogs, rabbits, 
guinea-pigs, and rats. Two donkeys inoculated have shown no 
reaction. The other, possibly identical with T. vivax found in the 
Kameroons, was fatal to cattle in five of the six cases in which it 
was observed; the other is still living. Sheep and goats have taken 
the disease; but in eight experiments six appear to have recovered, 
two only dying (in each instance small weakly animals, which also 
harboured intestinal worms). Dogs, rabbits, guinea-pigs, and rats 
appear refractory. One donkey inoculated has shown no reaction. 

4 (4) Experimental treatment was adopted in most of the 
naturally affected cattle, and it was eventually shown that atoxyl 
administered in 20 per cent, solution intravenously, in doses of two 
three and four grammes on successive days, caused the complete 
disappearance of trypanosomes from the blood; they however 



59 

recurred in from five to ten days, and further doses of atoxyl caused 
signs of poisoning. 

* According to the observations of the Liverpool workers, it is 
surmised that mercury in the form of corrosive sublimate will prevent 
the recurrence, when once the trypanosomes have disappeared from 
the general blood stream. It has been shown that cattle will tolerate 
the administration of one gramme of this salt given by the mouth, in 
I in 500 solution, on three successive days. It was necessary for me 
to leave Broken Hill before the combination of these two drugs could 
be tried in these doses, but the course of the experiments indicate 
that the greater the amount of mercury given, the longer the period 
of freedom from trypanosomes in the blood, and it may be shown 
that even more mercury than this could be tolerated by cattle, and 
would be necessary to bring about absolute absence of the germs 
and complete cure. 

4 (5) No exact experiments with regard to the mode of 
transmission of this disease were made. Two healthy cattle were 
walked through a narrow fly belt, and three tsetse flies (Glossina 
morsitans) were observed to bite. Fifteen days later both showed 
T. vivax in their blood and on the eighteenth day T. dimorphon . 
This is about the normal incubative period, and I have little doubt 
but that they derived the double infection at the time when the 
tsetse flies were seen to feed. 

‘ The investigations made into one outbreak, and all circumstances 
relating thereto, throw a very strong suspicion upon two species of 
biting flies as transmitters of this disease. These species belong to 
the genera Stomoxys (stinging house-fly) and Lyperosia, a very 
diminutive fly, similar in shape to the ordinary house fly. These two 
genera frequent farmyards, stables, and habitations, and breed in 
manure pits and dung. The former is incriminated in the natural 
spread of “ fly disease ” in Central South America and the Philippine 
Islands. 

‘ Another genus of biting fly which has become excessively 
prevalent as I leave here is the Tabanus , or “gad fly.” This fly is 
responsible for spreading “ fly disease ” in Algeria and India. 

‘ (6) My brief work and observations here have shown how urgent 
is the need for more research into this disease which is impeding the 
development of the country around rail head, and how imperative it 



6o 


is to continue experimental work on treatment, which shows with 
this combination of atoxyl and mercury indications of success which 
have never been obtained by any previous method of treatment. 

* It must be understood that these notes are merely for guidance, 
as it is impossible to make more dogmatic statements after so short 
a period of research, and that any deductions drawn therefrom are 
susceptible of modification/ 

It will be gathered from the preceding statement that the loss 
from disease is increasing—that they had verified its existence in 
36 cases; that they had isolated two distinct species of trypanosomes 
—one closely allied to T. dimorphort , the other possibly identical 
with T. vivax ; that large doses of atoxyl were beneficial; that with 
regard to the question of transmission, they walked healthy cattle 
through a narrow fly belt, and that three tsetse ( GL morsitans) were 
observed to bite. Fifteen days later T. vivax was found in their 
blood, and on the eighteenth day T. dimorphon ; further, after 
investigating an outbreak, they concluded that there was strong 
suspicion that two genera of biting flies acted as transmitters—one 
Stomoxys , the other Lyperosia. They also conclude that Tabanus 
may take a part in transmission, as it does in Algeria and India. 

The second communication is from Dr. Kinghom and is dated 
December 18, 1907, at Madona, and relates to the role of 
Gl. palpalis and GL morsitans respectively, in the transmission of 
sleeping sickness, and to certain practical measures for preventing 
the spread of sleeping sickness. 

1 Madona, December 18, 1907. 

' I left Madona on the 20th of November and went down the 
Luapula as far as Kazembi's, about two days south of Lake Mweru. 
From there I went East as far as Mshota’s and then returned to this 
place along the Luengo river. 

‘I did not look for GL palpalis along the Luapula particularly, 
as that had already been done, and I understand from Dr. Spillane 
that this fly exists along a large part of its course. However, I 
caught specimens of GL palpalis on the bridge over the Luongo 
and again opposite the Johnston Falls, both places on the main 
road from Madona to Kalungwisi. I also got specimens of 
GL morsitans in a village just at the commencement of the Falls 



6i 


After leaving Old Fort Rosebery, I did not see any fly along the 
whole of the rest of the route. 

* I found two cases of trypanosomiasis on the road to Kazembi’s. 
Both these had decidedly enlarged glands and trypanosomes were 
present in the gland juice. Both men had worked at Kambove. 
The finding of these cases is a verification of my earlier opinion that 
cases would be found scattered over the country among the natives 
who had worked in the Congo Free State. 

1 We have then, along the Luapula, all the factors which are 
necessary for an extension of the disease, i.e., cases of human 
trypanosomiasis and tsetse flies. Gl. pal pal is is found chiefly along 
the borders of the river, but also extends for a variable distance up 
some of the larger affluents. Gl. morsitans is found more or less 
plentifully over most of the country immediately behind the Luapula. 

1 There is an impression here that Gl. palpalis is the only species 
of tsetse fly which is capable of transmitting human trypanosomiasis. 
As you are aware, this question has not been settled decisively; on 
the contrary, from the results of work done in Uganda it seems clear 
that species other than Gl. palpalis can carry the disease and also 
that mechanical transmission is possible. While I do not believe that 
mechanical transmission alone is sufficient to account for the spread 
of the disease, it must not be forgotten that it is a possible means, 
and in the light of this I am afraid I must criticise the action which 
is being taken here. 

1 The Tanganyika Concessions, Limited, have here some six to 
eight thousand loads which have to be carried to Kambove. 
Permission has been given to recruit the labour for this in British 
territory. A strip along the Luapula, some ten miles in width, has 
been declared to be infected territory. If the natives required to 
carry these loads to Kambove are recruited in this “ infected strip,” 
they will leave here to proceed into uninfected territory on the Congo 
side of the river If they are recruited outside the infected area, 
they will have to come into this “ infected strip ” for the loads. In 
either case natives will be moving from uninfected to infected 
country, or vice versd. This, of course, is the one thing which should 
not be allowed. 

4 It is said that there are no Gl. palpalis on the route which is 
to be followed from here to Kambove. While this is the case, 



62 


GL morsitans is very plentiful, and the width of the road which is 
being cut (12 feet) is not by any means sufficient to banish them. 
I should strongly advise that the Tanganyika Concessions, Limited, 
be required to move the loads across the river and to find the carriers 
in the Congo Free State. I was informed by Mr. MacDonald, the 
agent of the Tanganyika Concessions, Limited, that food is very 
plentiful on the Belgian side of the river, and consequently natives 
also, so that there should be no difficulty in getting the carriers. 
However, this is a matter which does not intimately concern the 
Administration here, except in so far as a certain amount of money 
would be diverted from the country. It must be clearly understood, 
though, that if the disease is to be prevented from getting a foothold, 
all commercial considerations must give way until such time as the 
etiology and treatment of human trypanosomiasis are on a more 
satisfactory basis. 

‘ The other proposal that I feel is unjustified is one to establish 
a temporary segregation camp near Madona for the cases now known 
to exist. It is impossible to have this camp more than four or five 
miles distant from this station, as otherwise it would be too far away 
for easy access, and since natives are returning from Kambove, a 
medical officer is required here to examine them. The only place 
available within this distance, with a suitable water supply, is on the 
Luafumu river. Just around here, however, Gl. morsitans is fairly 
plentiful, so that there exists a risk of the disease spreading, however 
minimised by extensive clearing. To digress for a moment, I have 
a specimen of GL palpalis caught on the verandah of the assistant 
magistrate here, although the whole front of the river has been 
cleared for some distance up and down from his house (the house lies 
200 yards from the river bank, which has been absolutely cleared for 
400 yards in all, and beyond this cleared for gardens). Mr. Hughes, 
the assistant magistrate, informs me that there are places in the 
Luapula division eminently suited for a segregation camp, i.e., there 
are no tsetse flies, no villages and plenty of water. Of course until 
more medical men are available, some such course (i.e. a temporary 
segregation camp) will have to be adopted if the cases are to be 
treated at all, but the British South Africa Company should take 
steps to increase its medical staff immediately, so that the situation 
may be met energetically. 



63 


‘ In order to avoid misunderstandings, it should be clearly defined 
that the matter is one for which the medical officer is responsible 
and not the native commissioner. There is some tendency to reverse 
this order of things, but this should not be. The situation should be 
entirely under the control of the medical officer, and any assistance 
granted him from the native department should be subject to his 
direction. 

‘ I was informed by Mr. Beringer, whom I met at Kazembi’s, that 
the natives are crossing the northern frontier very freely. There are 
a lot of Swahili traders there engaged in smuggling, and these 
encourage native movement. An ordinance has been passed which 
will require these to leave the country. This of course is in 
accordance with the advice given by Todd some time ago, and was a 
very necessary proceeding. If it has not been done, I think some 
legislation should be passed which will enable the officials in charge 
of the sleeping sickness work to deal with any European who may 
feel disposed to ignore the regulations which have been made 
regarding the crossing of the borders with natives. So far as is 
known here, there is no penalty attached to this, so that if a European 
should go, say from Madona to Broken Hill through the Congo Free 
State, there is no means of dealing with him. In several cases special 
permission has been given for this to be done, and as the natives 
were recruited in the infected area, a dangerous precedent has been 
created. The Administration of North-Western Rhodesia should 
consider the advisability of forbidding the entrance of any native 
from this infected territory into its country. 

1 The whole question may be put thus—As a result of the finding 
of Gl. palpalis and cases of human trypanosomiasis along the 
Luapula river, a strip ten miles wide bordering the stream has been 
declared infected territory and all movement from it stopped. Yet 
it is proposed to allow the Tanganyika Concessions, Limited, to 
move some six thousand odd loads with carriers from North-East 
Rhodesia through country abounding in tsetse flies to a district from 
which the cases now present in the country have come. In addition 
natives have been allowed to proceed from the infected territory 
through non-infected parts of the Congo Free State and North- 
Western Rhodesia. In view of what we know at present of the 
etiology of sleeping sickness, I think that this is a mistaken policy. 
I would urge the adoption of the following suggestions: — 



6 4 


1 i. The Luapula should be closed absolutely until it has been 
proved definitely that GL morsitans cannot spread human trypano¬ 
somiasis. In my last letter I suggested how the river might be closed. 

‘ 2. Legislation should be provided whereby any person taking 
natives across the border can be prosecuted and fined. 

‘ 3. The Tanganyika Concessions, Limited, should be required 
to find carriers for their loads in the Congo Free State. 

‘ 4. The Administration of North-Western Rhodesia should 
forbid the entrance of any native into its territories from any infected 
area in North-Eastern Rhodesia. 

* 5 * Segregation camps for patients should be stationed in a 
district free from tsetse flies of any species. In order to do this the 
British South Africa Company will have to increase its medical staff. 

‘ 6. The medical officer should be given charge of the work and 
afforded such assistance from the native department as may be 
necessary to carry out his instructions. 

‘ As I have said before, I do not think that the disease has any 
hold on the country as yet, and if the matter is faced at once 
regardless of any other considerations, it can be prevented from 
spreading to any extent. Otherwise the danger of this is great, so 
far as our present knowledge extends. 

‘ Yours faithfully, 

‘(Signed) Allan Kinghorn.’ 

In a letter dated Madona, January 28, 1908, to Dr. Barratt, of 
the Expedition which was sent out to investigate Black water Fever 
in Nyassaland, and if practicable to act in concert with the Sleeping 
Sickness Expedition, Dr. Kinghorn makes a statement in which he 
emphasises the importance of regarding all biting flies with suspicion, 
and states that every effort should be made to find exactly what flies, 
tsetse or others, are capable of carrying the virus. 

‘ As regards “ fly,” GL palpalis has been found on both sides of 
the Luapula from Kapwepi’s to Kasiwa’s, a village about two days 
south of Lake Mweru (by Dr. Spillane and myself). It has also been 
found by Dr. Spillane along the whole of the British coast of Mweru, 
around the south end of Lake Tanganyika, and also along parts of 
the larger affluents of the Luapula and Kweru, notably the Mansa 
and the Kalungwisi. I found it on the Luongo a short distance from 
its mouth. From Kasiwa’s to Mweru, on the British side, the Luapula 



65 


is bordered by swampy plains some miles in width, so that the 
conditions are unfavourable for this fly. On the Belgian side, this 
does not obtain to such an extent, so that the fly is probably present, 
although I have no definite knowledge as to this. GL morsitans , 
and possibly the closely related pallidipes and longipalpis> is widely 
spread over the country, however. On the road from Broken Hill 
to Fort Jameson, I found them at one spot in the Machinga Hills, 
from the base of these right across the Luanga to within a day of 
Petauke, and near the Sasare mine. From Fort Jameson to the 
Luapula, they occur from the Luangwa to the base of the Machingas. 
There is a sudden rise here to the plateau of 2,500 feet or so, and on 
top of the hills I did not see fly again until I got to the Kasanka 
river. In this vicinity they are very plentiful, and stretch over to the 
Luapula and up north past Chitambo's through the country to the 
east of Bangweulu. They may be said to be found in most of the 
Luapula division. 

‘ I have found three cases of sleeping sickness in the course of 
my work, all with a history of having worked in the Katanga mines. 
All had markedly enlarged glands, and, at the time I saw them, 
appeared perfectly healthy. Fly (GL morsitans) was present in the 
village of at least one case. Considering the thousands from this 
country who have worked in the Katanga mines, I am perfectly 
convinced that there are many more cases scattered through North- 
East Rhodesia, as the labour was not drawn from any one particular 
district. 

1 As you are aware, all the work which has been done goes to 
show that the transmission is mechanical. While I think this is not 
sufficient to account for the rapid spread of the disease in view of 
the great difficulty experienced in getting positive results, it is a fact 
which cannot be too strongly emphasised. There is too great a 
tendency to regard Gl. palpalis as the only infecting agent. That 
this is not so has been shown by the work in Uganda, where successful 
transmission experiments were made with GL fusca. Again the work 
on cattle trypanosomes shows that while one or two species are 
normally concerned in the extension of the disease, it is quite possible 
to effect this by the use of not only other species of tsetses but also 
other distinct genera, e.g. Stomoxys and Tabanus. This surely is 
sufficient to demonstrate that, at present, all biting flies, and 



66 


particularly all species of Glossina , must be regarded with suspicion. 
While, perhaps, the task of controlling the spread of sleeping sickness 
would become impossible if every biting insect had to be considered, 
it is not so with regard to the tsetse flies, and I would hope that on 
every occasion on which you have the opportunity you would 
emphasise the importance of not simply regarding GL pal pal is alone 
as dangerous. The question of the etiology of the disease is in a 
more unsatisfactory condition than the treatment, at all events from 
the prophylactic point of view, and it is most important that some 
definite effort should be made to find exactly what flies, tsetse and 
others, are capable of carrying the virus. 

‘ As to enlarged glands, I have found that a fairly large percentage 
of the natives (roughly 30 to 40 per cent.) have palpable glands which 

come chiefly under the “ +-” group of Dutton’s and Todd’s 

classification, though there are also a number of “ H-” glands. On 

puncture these were found to be uniformly negative. This occurrence 
of enlarged glands rather complicated the diagnosis, for it means that 
the statement " every negro with enlarged glands must be considered, 
until the contrary is shown, to be a case of trypanosomiasis,” cannot 
be accepted for North-East Rhodesia, and, presumably, for Nyassa- 
land as well. Consequently a trained medical officer is the only 
person who can look for the disease satisfactorily, since puncture is 
necessary. I have advised that the British South Africa Company 
should have special medical officers to travel through the country 
looking for cases and getting them isolated at once. Whether it 
will be necessary for like measures to be taken in Nyassaland is 
uncertain as yet. 

* The chief danger for the Protectorate lies in the spread of the 
disease to the north end of Lake Nyassa. Trypanosomiasis is 
endemic on Tanganyika at Vua in the Free State, a short distance 
above the Congo-Rhodesia border, and between the people on either 
side of this there is unrestricted communication. Gl. palpalis is 
found right round the southern end of Tanganyika, so that it is 
extremely probable that cases of the disease are present in that part 
of the country. Whether the disease is to be found in German East 
Africa in this neighbourhood is not known. At the north end of 
Nyassa GL fusca has been found, and this, we now know, can carry 
the disease. If once a case gets over where these flies occur an 



67 


epidemic might easily be started. I believe natives of the 
Protectorate are carrying loads from Karonga to Kasana, and as large 
numbers of natives from the Awemba district of North-East Rhodesia 
have worked in the Katanga, cases probably exist amongst them, 
and the possibility of their infecting negroes from Nyassaland 
must not be overlooked. It therefore seems to me that the north 
end of Nyassa is the part which requires most attention from the 
authorities of the Protectorate. It would also be well to bear in 
mind that natives of the Protectorate have been working in the 
Katanga and some are only returning home now. Some of these 
may be infected and, in fact, I should not be at all surprised to hear 
at any time that cases of human trypanosomiasis had been found in 
Nyassaland. 

4 As soon as we get carriers we intend to travel across to the 
north end of Nyassa in order to examine the conditions there 
personally. We should be there in four or five months at the outside. 

4 Yours sincerely, 

‘(Signed) Allan Kinghorn.’ 

The members of the Sleeping Sickness Expedition, having seen 
in the 4 Times ’ of 13/12/07 an article dealing with Game Preservation 
and its relation to Ngana, make the following statement : — 

‘The Preservation of South African Wild Game. 

4 It may not be considered out of place for us to enter into the 
discussion on the subject which appeared under the above heading 
in the issue of the “Weekly Times” for December 13, 1907, as 
the article bears most directly upon the question of tsetse flies and 
the disease they transmit which we were dispatched by the Liverpool 
School of Tropical Medicine in the spring of last year to investigate 
on behalf of the respective Governments of British territory, North 
of the Zambesi. 

4 We hold no brief, and are actuated by no party: we merely 
discuss the problem from the scientific standpoint, and if sentiment 
sways us towards the prevention of vandalism in the works of man 
or nature, there is not lacking evidence which completes your 
correspondent’s propositions and shows the danger existing. We do 
not presume to enter the lists on a footing with such observers as 



68 


Sir Alfred Sharpe, F. C. Selous, F. J. Jackson, or R. G. Harger, 
whose intimate knowledge of their respective countries and long 
residence there places them in a position to have personally watched 
the passage of events. We arrive as observers of the question as it 
stands to-day in the parts of the Continent over which we have 
travelled. 

* The problem may be stated in the following paragraphs which 
appeared in the article mentioned:— 

* “ I. ... That certain species of tsetse fly exist only where 
wild game exists. 

‘ “ 2. That certain germs exist in the blood of wild animals, 
which .... when transferred by the tsetse .... are the cause 
of Ngana or tsetse fly disease. 

‘ “ 3. If therefore the propositions quoted (above) can be 
proved valid the game must be destroyed ; there is no help 
for it.” 

‘ 1. We have the widespread and uncontroverted statement that 
in many districts of South Africa the tsetse flies have disappeared; 
the reasons advanced being (1) the progress of civilisation, which 
implies the opening up of the country, the presence of men and guns, 
and the destruction of some game, the flight or retreat of those 
remaining: and (2) the epidemic of Rinderpest which swept over the 
country towards the middle of the last decade, killing off buffalo and 
antelope, and so removing the only food upon which the tsetse was 
held to exist. 

‘ North of the Zambesi a precisely parallel state of affairs has 
obtained. Rinderpest raged, killing cattle in front and leaving the 
veldt strewn with carcases of game. A railway has been built, mines 
and mining centres opened up, and farms and mission stations are 
scattered over the territory, each and all implying the advent of men 
and guns, the destruction and the driving back of game. But the 
results have not been the same : wherever there is evidence that 
tsetse existed in the past, there it may be found to-day. Naturally 
we do not speak of the centre of a busy mining camp: the retreat 
of fly from this would be as from a native village; it is a relative and 
not an absolute disappearance. Further, there is much evidence that 
the tsetse now holds sway over roads and areas where they were 
unknown a few years ago. 



6 q 


‘ We are told that the tsetse of South Africa depend upon game 
for a living: it is true that science does not yet know for certain that 
this fly can obtain a living on anything but blood. Many other biting 
flies can maintain a vegetarian existence for a time at least, and the 
circumstantial evidence is certainly strong that the genus Glossina 
can do the same. In what other way can be explained the not 
infrequent occurrence of districts uninhabited by man and in which 
game, spoor or even the ordinary indications which go to make a 
“ game country ” are wanting, yet in which tsetse abound in their 
thousands, congregating around the traveller and his carriers in 
numbers impossible to ward off P An estimation of the odds against 
any one of these flies obtaining a meal of mammalian blood in the 
course of a year would indeed be work for a statistician. Further, 
there are districts and large areas wherein game of all local varieties 
is plentiful and in which no tsetse can be found. What explanation 
can be offered? Only that, like any other being, the members of 
the genus Glossina have their likes and dislikes, and select for a 
home a locality suitable according to their tastes. Even amongst the 
members there is individualism, for one species— Gl. palpalis —will 
not live away from the immediate vicinity of water; and only such 
water, too, as is open, and is bounded by defined banks which carry 
a sufficiency of timber or scrub to provide him with the necessary 
amount of shade. As every traveller in Northern Rhodesia knows, 
the tsetse fly of that country— Gl. morsitans —does not make a point 
of these conditions; but it does insist on shade, preferably that 
furnished by the half-grown trees which constitute the virgin bush, 
the ground between which is covered with grass, smaller trees or 
scrub. It does not live on an open flat, dambo or vlei, and it is 
quite patent that the presence of water is secondary in its estimation. 
From this it is obvious that there is much truth in Mr. Dunbar’s 
suggestion that local variations in the tsetses may account for some of 
the discrepancies in the accounts given by various writers, for he, in 
turn, introduces another species of tsetse into the argument. 
Gl. fusca, the large species to which he refers, has been shown to 
exist at both the North and South ends of Lake Nyassa; its haunts 
and habits are not well known, but there is some indication that it 
favours similar country to GL palpalis , i.e., water and shady banks. 
As some species of game undoubtedly confine themselves to certain 
F 



?o 


types of country, and as the tsetse is also shown capable of selective 
powers, may it not be that the association of fly—used without 
distinction of species—and particular varieties of game is but a 
natural inclination of both beings to the same locality, and not a 
peculiar affinity between the individuals? 

‘ These statements do not answer why the tsetse (believed to have 
been both Gl. morsitans and its relative Gl. pallidipes) has 
disappeared from parts of Africa South of the Zambesi, but they go 
to show that game destruction by civilisation and rinderpest is not the 
only factor concerned. Until we are better acquainted with the 
bionomics of the genus Glossina , by observation and experiment 
conducted on scientific lines, the statement “ that certain species of 
tsetse exist only where wild game exists ” must remain controversial, 
but with a bias to the negative, if, indeed, it must not be negatived. 

‘ 2. The second of your correspondent’s propositions may be 
dealt with in a more dogmatic fashion. “ . . . . that certain germs 
(trypanosomes) exist in the blood of wild animals which produce in 
them little or no ill effect; and that these germs when transferred by 
the tsetse . . . are the cause of Ngana or tsetse fly disease.” 

‘ The trypanosomes of various countries all possess features in 
common with those of Zululand, which was shown by Bruce to be 
incapable of living outside an animal body, and to be spread by a 
biting fly, to the exclusion of all other natural means. They differ 
only in minor structural points, but especially in their actions upon 
stock. From all, death is the result in a susceptible animal, but 
towards some trypanosomes, cattle, towards others, camels, possess a 
degree of resistance approaching or equalling that enjoyed by wild 
game towards the Zululand form. 

* As the germ cannot live in air, forage or water, it can only be 
derived from a pre-existing case; a reservoir, which like big game 
may not suffer from the disease, or an animal already sick. Bruce 
showed that buffalo, koodoo, wilderbeeste, bush-buck and hyaena may 
be that reservoir in Natal; Dutton and Todd proved that the bush- 
buck in the Congo Free State harboured the trypanosome of fly 
disease there; and in North-Western Rhodesia we have found the 
germ in bush-buck and Lichtenstein’s hartebeest. This germ is not 
the same as that found by Bruce, but the sickness in cattle is 
identical. In North-Western Rhodesia, and when properly studied 



7i 


the same will probably obtain elsewhere, there is more than one 
species of Trypanosoma , and though the effects towards some 
animals—cattle—are the same, the action on others—goats and sheep 
—is different. May this, too, not account for many of the 
discrepancies in travellers’ reports? For example, the opinions 
regarding the susceptibility of the donkey are very diverse; we found 
that this animal took one of the Rhodesian trypanosomes, whilst a 
second donkey did not become affected with the other form of the 
germ which occurs there. 

‘ In many countries, and districts of countries too, where trypano¬ 
somiasis is rife, the amount of game is negligible. From whence 
does the virus, the causa causans , then arise ? When the conditions 
are examined it is found that there is always at least one species of 
domestic animal which is more resistant than another: it may never 
be sick; or if it does take the disease it may live for two or three 
years like the camel with the Indian fly disease, and it may even 
recover. In South America, fly disease is seen chiefly in the horse, 
and cattle rarely suffer, but they, in common with the camel of India, 
take the place of the non-existent game, and act as a reservoir which 
under the required condition can issue supplies of trypanosome to 
all comers; causing death in those susceptible and converting the 
others into additional reservoirs. 

‘ But what is the required condition without which the presence 
of a reservoir is not greatly to be feared? Bruce showed it to be a 
biting fly. In Africa travellers blamed the tsetse; in India natives 
incriminate the “horse fly” ( Tabanidae ), known in Rhodesia 
as the “ hippo fly,” and a smaller variety locally called “ blind fly ” 
(genus Haematopota') ; in South America and parts of Asia the 
“ stinging house fly ” (genus S(omoxys) is blamed. 

‘ With few exceptions no one in South or Central Africa has even 
suggested the possibility of any fly other than tsetse causing the 
disease; and since the time when Bruce proved the connection 
scientifically, what was at first a mere suspicion influenced by native 
tales has become a conviction, to the exclusion of all other flies. It 
is not necessary to look far for the reason. Stock owners knew they 
could keep their animals around the farmstead and in certain areas 
with impunity, even though horse flies and the stinging house flies 
were present. Compared with the tsetse these other flies are 



72 


“ domestic ” ; they are limited in distribution on the veldt; their 
haunts and habits keep them from the free association with game 
that tsetse enjoy in the zones of which they and game are common 
residents; and they have not a fraction of the voracious nature and 
pertinacity possessed by the Glossina . On frimd facie grounds 
they cannot be incriminated to the same extent as those which live 
among the reservoir and which will follow an ox for miles in the 
hope of obtaining a meal. It is this very “ domestic ” nature of the 
other flies, however, which renders them accessories par excellence in 
spreading the disease when a domestic animal, controllable and 
housed, is the reservoir. Let the farmer in a tsetse fly country expose 
one of his animals until it acquires the disease and then herd it with 
the rest in a kraal inhabited by Stomoxys or other biting flies such as 
Lyperosia or Haematobia. The reservoir and the biting fly are 
present in the midst of healthy and susceptible stock; and the results 
have been severe enough to be well appreciated by some agriculturists 
in Northern Rhodesia. 

1 3. This second proposition is proved valid: Buffalo, Koodoo, 
Wildebeeste, Bush-buck, Hartebeest and Hyaena have been shown 
“ accessories before the fact,” and if we adopt your correspondent’s 
view, they must be destroyed. 

‘ We enter a plea that sentence may not yet be passed. To quote 
from the article : —“ A hasty sentence of wholesale death is one whose 
execution, once accomplished, can never be annulled.” Every month 
is adding to our knowledge concerning the disease in all parts of the 
world, and the rapid accumulation of evidence all goes to show that 
efficient sanitary police measures adapted to the requirements of each 
district, and even to each individual owner, formulated and carried 
into effect by an adequate staff of veterinary officers will do much 
to control trypanosomiasis in the domestic animals. There is no 
proof that the eradication of the game will be followed by the 
disappearance of fly disease: South Africa cannot be accepted in 
evidence, for the factors said to have been causative there have been 
without effect in other parts of the Continent. We have indicated 
that the destruction of the tsetse would not prevent the transmission 
of the germ under the conditions which obtain in countries where 
both game and Glossina are non-existent; and though it is patent 
that the extermination of the reservoir would, d priori , check the 



73 


disease, vve have no proof that in Central Africa a storehouse for the 
germ other than game may not exist. 

‘ Irrevocable measures are not called for to satisfy the materialistic 
and often speculative desires of the few: for the many, a proper 
knowledge of the disease and the adoption of trained advice as to 
how it can often be avoided, may tide over the time till an effectual 
cure or a specific means of prevention be made known; and we 
earnestly urge that less draconic measures may be given a full and 
fair trial before the death warrant of the greatest natural attraction 
of Central Africa be signed. 

R. Eustace Montgomery. 
Allan Kinghorn. 

Madona, North-Eastern Rhodesia, January 28 th y 1908. 

In correspondence dated Madona, February 7th, 1908, the investi¬ 
gators state: — 

‘ With regard to the report we should like to remark that we 
disagree with the policy of the Tanganyika Concessions, Limited, in 
sending loads to Kambove with natives fr.om this country. By 
refusing permission to recruit labour for the mines, and still allowing 
loads to be taken in, we think that the Administration is placing itself 
in rather a false position. The Tanganyika Concessions, Limited, 
if required, would, we think, be able to find carriers in the Katanga 
as they have to do with regard to loads coming in via Broken Hill. 
They say they are taking precautions to prevent these carriers becoming 
infected, but these precautions only consist in cutting a road about 12 
feet wide along a route in which 67 . morsitans is very plentiful, and 
by the establishment of food stations away from the villages. The 
natural tendency would be for the natives to try and stay in the 
Katanga to work, and in at least one instance a negro has run away 
from the gang when returning to Madona. Of course, he may not 
have gone back to the mines, but this is the likeliest thing, as they 
are paid on getting back there. 

4 Another rather anomalous proceeding of the local government is 
the official declaration of the 10 mile strip along the Luapula to be 
infected, so that natives come into this area on their way to Kambove, 
which is 44 uninfected.” Thus we have the passage of negroes from 
infected to uninfected territory and vice versd. 

4 Yours faithfully, 

4 Allan Kinghorn. 



74 


‘ Madona, February 4 th, 1908. 

‘ Dear Dr. Hearsey, 

‘ In a recent number of the “ British Medical Journal ” I notice a 
reference to your last year’s report, and as you consider the question 
of sleeping sickness, I should like to get a copy of it, and also of the 
map showing the distribution of tsetse flies, if you will kindly send 
them to Abercorn. 

‘ I notice also that the road from Tanganyika to Nyassa is now 
unused, and that Dr. Todd’s prediction as to the infection of the 
South end of the last-named lake has not been fulfilled Unfortu¬ 
nately this is not the case. Evidence is at hand that cases now exist 
there, and as Gl. pal pal is is found along its shores, the danger of 
a wide dissemination of the disease is great. It is not surprising that 
this is true, for although fly was known to exist along the Belgian 
side of the lake, certainly as far South as Vua, and also that imported 
cases were present at Moliro (1901) and Baudoinville (1902), nothing 
was done by the Congo authorities to check the spread of the disease, 
and only last year the natural consequences of this were shown by a 
report that the disease had become endemic at Vua. As natives on 
either side of the border have been communicating uninterruptedly, 
it would be a matter for some surprise if cases had not Deen brought 
into British territory. In German East Africa cases have been 
present at Ujiji, but whether there is any danger of the disease 
spreading southwards to the north end of Nyassa I cannot say. GL 
palpalis is found on the German portion of Tanganyika, certainly 
below Bismarckburg. 

‘ Along the Western border here I have found three cases of try¬ 
panosomiasis, all with a history of having worked in the Katanga. 
I consider that these cases originated there without doubt. No 
indigenous cases have been found here, and as GL palpalis occurs 
along the whole of the Luapula from Kapwepwi’s to Kasiwa’s (two 
days south of Mweru) and the people are on the river constantly, 
these would have been expected had the disease been indigenous. 

‘ As a result of the extensive importation of labour into the 
Katanga, cases of sleeping sickness will probably be found in all the 
districts from which it was drawn. This applies not only to 
Rhodesia but to Nyassaland as well. I am not aware how many have 
gone from Nyassaland, or whether they have been watched since 



75 


returning, but whether or not, there is any danger of the disease 
coming in from the North, this route must not be forgotten. Skilled 
workmen and raw labour as well, from Nyassaland, are still in the 
Katanga. The presence in the Protectorate of 67 . fusca , known to 
be capable of transmitting T. gambiense , is of importance if cases are 
found. 

‘ I cannot deprecate too strongly the continued permission to the 
Tanganyika Concessions to carry loads from Madona to Kambove 
with Rhodesian natives. The anomaly of this is at once apparent 
when it is remembered that free recruiting for the mines has been 
stopped. Another anomaly is that while Kambove is not regarded 
as being in the “ infected ” area of the Katanga, a strip io miles wide, 
along this side of the Luapula, has officially been declared to be 
infected. By allowing natives to pass, therefore, from an infected to 
a non-infected district the extension of the disease is inevitable. In 
my opinion the Luapula should be absolutely closed. 

‘ There is a possibility of slight danger being attached to the 
Karonga-Kasama route. Cases of sleeping sickness will probably be 
found in the Awemba district of North-East Rhodesia, and tsetse-fly 
exists along the road in places. At present, however, I am inclined 
to think the danger is remote. 

1 When I was in Fort Jameson, Dr. Barclay told me that the 
results of the palpation of all the neck glands, not simply those in 
the posterior triangle, had been included in the report sent to the 
Colonial Office early in January (dated Zomba, January 24th, 1907). 
If this is correct for all the figures, they would need to be corrected. 
Were any of the “ 4- ” cases punctured ? In view of what we have 
found in Rhodesia, this would be advisable. The cases I found had 
+ ” glands, and, of course, showed the parasite (of five “ + ” cases 
seen, three were infected). Of 9,005 natives examined by me 
between July and December, 20*85 % had palpable glands, mostly 
very few in number and shotty to the touch. 

‘ One other point on which I should like to touch is the question 
of what tsetse flies are able to transmit the disease. As you will 
know Gl. pal pal is and Gl. fusca have been shown to be capable of 
doing so. Whether or no the other species can effect the transmission 
as well, is not known, but since mechanical transmission is the only 
method of which we have any knowledge at present, the part these 



7 6 


may play, given the presence of cases, becomes an important consi¬ 
deration. Even though it should eventually be shown that the 
trypanosomes have a life cycle in any one tsetse fly this mechanical 
transmission cannot be absolutely ignored 

4 Yours faithfully, 

‘Allan Kinghorn. 


‘ The Principal Medical Officer, 

1 Zomba, Nyassaland.’ 



77 


THE INCIDENCE AND PROPHYLAXIS OF 
HUMAN TRYPANOSOMIASIS IN NORTH 
EASTERN RHODESIA 

BY 

ALLAN KINGHORN, M.B., Toronto, 

JOHNSTON COLONIAL FELLOW, UNIVERSITY OF LIVERPOOL 
AND 

R. EUSTACE MONTGOMERY, M.R.CV.S. 

(First interim report of the Expedition of the Liverpool School of 
Tropical Medicine to the Zanibesi i 190J-08) 

When the Expedition to the Congo returned to England in the 
autumn of 1905, Dr. Todd stated that there was reason to fear an 
extension of human trypanosomiasis from the Congo Free State into 
British territory. This statement was based principally on the facts 
that the disease was spreading steadily southwards, that imported 
cases of sleeping sickness existed practically on the borders of 
Rhodesia, at Moliro and Baudoinville on Lake Tanganyika, and that 
one of the main trade routes ran from this lake to Nyassa. The history 
of the spread of the disease in the Congo showed that it had been 
carried from point to point along the main caravan roads, often with 
startling rapidity. 

In the following year, reports were received that an imported case 
existed at the Belgian post of Kasenga on the Luapula River, and in 
September, 1906, Dr. Todd and one of us were informed by Mr. F. W. 
Amott, of the Garanganza Mission, that cases were present at Lukafu 
on the Lufira River. In November, Dr. Todd received from 
Dr. Massey, then of the Tanganyika Concessions, Limited, a 
communication stating that the disease was endemic in the villages 
around Lake Kisale and some of its confluents, and further, that about 
9 per cent, of the carriers from Kabinda to Ruwe were infected 
(When Dr. Todd went through this part of the Congo, he found that 
about 13 per cent, of the people in the neighbourhood of Kabinda 
were infected, but that fish sellers from the South, whom he saw there, 
were free from the disease.) In about a year and a half then, June 



78 


1905 to November 1906, the disease had become endemic around 
Lake Kisale, having been imported from Kabinda. As loads were 
constantly being brought to the Katanga mines from these infected 
areas, the greatest danger existed that foci of the disease would be 
established there, especially as Dr. Massey had found tsetse flies 
( 67 . palpalis and Gl. morsitans ) in the immediate vicinity of some of 
them. Most of the labour for these mines had been recruited in 
British territory, so that the gravest fears were entertained that the 
disease would be carried into Rhodesia by some of these men on 
returning to their homes. Finally, about the same time a report 
reached home that the disease was invading the Western shores of 
Lake Mweru. 

In consequence of this knowledge, representations were made to 
the Colonial Office by the Authorities of the School, pointing out the 
danger of trypanosomiasis being carried into British territory, and 
asking for support for an expedition which it was proposed to send 
to the threatened districts to study more fully the local conditions. 
At the same time the British South Africa Company were asked to 
assist, as North-Eastern Rhodesia would probably be the first to 
suffer. This proposition was eventually accepted, and the present 
expedition sent out. Broken Hill was reached on June 16, 1907, 
and shortly afterwards one of us left for Fort Jameson. 

The following report, it will be understood, does not discuss the 
situation fully, as we have seen but a small portion of the country. It 
is believed, however, that enough evidence has been accumulated to 
judge of the probable occurrence of human trypanosomiasis, and also 
to judge the value of the various measures which have been suggested 
for combating the disease. 


ROUTE 

On the accompanying map the route followed to date will be 
found, and also those areas of North-Eastern Rhodesia in which 
species of Glossina are known to occur. This information, of course, 
is not complete at present. So far as possible the main roads were 
followed, as it is along these that experience has taught us the 
disease is usually carried. As we went from village to village, all the 
inhabitants were palpated, and in those cases in which it was possible, 
gland puncture was performed. This enabled us to control the value 



79 


of gland palpation as a means of diagnosis so far.as a country was 
concerned in which the disease was supposed to be non-existent. 
Most attention has been devoted to the Luapula division, as large 
numbers of natives from this district have worked in the Katanga, and 
as, in the natural course of events, it would be the first portion to be 
invaded. The other area from which the disease was expected to enter 
Rhodesia was along the border between Mweru and Tanganyika. As 
yet we have not been able to visit this part, but evidence is 
accumulating that the prediction made by Dr. Todd is in course of 
fulfilment. 


DISTRIBUTION OF TSETSE FLIES 

A. In North-Eastern Rhodesia. 

i. Glossina pal pal is. In brief, we now know that this species 
exists along a large part of the Luapula and some of its confluents; 
on the shores of Lake Mweru; and around the southern end of 
Lake Tanganyika. 

It was first found in 1906 by Dr. Noble at Kasenga and the 
Nafunta Falls on the Congo side of the Luapula. This year it was 
found by Dr. Spillane along this river from Kapwepwi’s to Kasiwa’s 
village, and along the British portion of the two lakes. We have 
been able to confirm these observations as regards the Luapula. It 
was also found by Dr. Spillane for some distance up the Mansa and 
Kalungwisi rivers, and we have found it up the Luongo a short 
distance from its mouth. 

Along the portion of the river mentioned above, the bush extends 
right to the margin of the water, where it assumes a more luxuriant 
growth and affords abundant shade. From Chongola’s to Sakontwi, 
the river is bordered by wide flats almost completely destitute of 
vegetation. From Sakontwi to Kapwepwi’s the shore is fringed by a 
single, more or less continuous, row of bushes and small trees which 
project obliquely outward over the water. Behind this there is a 
bare, treeless strip of ground varying in width from 25 to 200 yards, 
and beyond this again the ordinary thin bush of the country begins. 
This lack of continuous shade behind the river and the small amount 
of shelter afforded by the fringe of bushes probably account for the 
absence of Gl. pal pal is. From Kasiwa’s to Mweru, the conditions are 



8o 


somewhat similar except that swamps, one to five miles wide, replace 
the dambos. This fly, 67 . pal pal is , is most readily found by paddling 
just along the trees which line the shore, and success often follows 
this manoeuvre when a search on the shore itself has been fruitless. 
As a rule, the fly will not come out to the canoe if very far from the 
shore, although occasionally we have noticed it at a distance of from 
ioo to 150 yards. As has been observed before, we have found it to 
be most active in the middle portion of the day when the sun is 
shining brightly. Early in the morning and late in the evening are 
unfavourable hours to look for it. We have also been able to confirm 
the observation made in Uganda 1 that the fly is absent from those 
portions of* the shore where the trees are replaced by grass and 
water weeds for any distance. 

2. Glossina morsitans . While we use this specific name, we wish 
it to be understood that the identification of our specimens is not 
complete, so that in addition to Gl. morsitans proper, the closely 
related pallidipes may exist. This will be discussed more fully at a 
later date. 

This fly has an extremely wide distribution, not in definitely 
defined belts, such as are said to be found farther South, but more 
or less generally scattered over the whole country. For the purposes 
of this report it is not necessary to give all the situations in detail: 
it will suffice to say that it is found in most of the districts in greater 
or lesser numbers. While these areas are marked on the map, it 
must not be concluded as yet that they are not present in the 
unmarked portions. 

Like Gl. pal pal is, we have found them to increase in activity as 
the sun approaches the zenith, and to be fewer in number and less 
voracious on dull days. Their activity is maintained well on towards 
sunset, and we have noticed an occasional one flying into the tent 
after this time. Water and shade are not such necessary factors to 
this fly as to pal pal is. We have observed it at least three miles from 
water, and have been informed by one of the officials that he has seen 
it ten miles from water. The amount of shade afforded by the thin 
type of bush peculiar to the country is very small, in the dry season 
practically none. A certain amount of shelter is required, however, as 
this fly is not found on the open dambos which break the continuity 
of the bush. 



8i 


The consensus of opinion throughout Rhodesia is that tsetse flies 
are steadily increasing in numbers, regions which only a few years 
ago were free being now heavily infested by species of the genus, 
i.e., with Glossina morsitans and possibly pallidipes, This increase 
cannot be ascribed to a corresponding increase in game, since large 
areas exist where game is practically absent and tsetse flies abound, 
while in other parts the reverse holds good. 

B. In the Nyassaland Protectorate. 

Glossina palpalis has not been found here, but other species are 
known. Gl. fusca has been observed in the Karonga district, and 
again in the neighbourhood of Chiromo. Gl. pallidipes was found 
in the former area, and Gl. morsitans is present in various parts of 
the country. 

C. In the Katanga District of the Congo Free State. 

1. Glossina palpalis. From Dr. Massey we learn that this fly 
occurs on the West shore of Mweru; at the junction of the Dilukwe 
and Lufira rivers ; at Nkoni Hill on the Lufira ; on the Lukulegi river 
near the Congo-Zambesi watershed ; at Busanga, junction of the 
Lufupa and Lualaba (tin mines); on the South Kaluli, at the cut 
road crossing from Ruwe to Mazanguli’s; on the Lualaba, from the 
Kalenga Falls to Chisamba; on the Inje river, running into Lake 
Kisale. 

2. Glossina morsitans. In the whole region bounded by the 
Lualaba and Lufira; along the road from Kambove to Madona; in 
the neighbourhood of Busanga; on the Lubudi river, and at its 
junction with the Mkuleshi; on a line drawn from Ruwe to 
Kansanshi, in two places. 

The disease is endemic in this part of the Congo along the 
Lualaba, from its junction with the Lubudi to its point of exit from 
Lake Kisale, and on the Lufira around the Government post of 
Kayumba and at its junction with the Dilukwe. Moreover, the whole 
of the Lufira from Kisale to Mwenda’s village, a short distance from 
the Nkoni Hill Mission and the local administrative post of Lukafu, 
is suspected. We have also mentioned above that imported cases of 
the disease were present at Nkoni Hill in September, 1906. 



82 


GLAND PALPATION AND PUNCTURE 

Since Greig and Gray 2 noted that trypanosomes were fairly 
constantly present in the enlarged glands of persons suffering from 
trypanosomiasis, the efficiency of this means of diagnosing the disease 
in its earlier stages has been repeatedly demonstrated. Dutton and 
Todd 3 were the first to recognise its practical importance, and their 
experience led them to make the statement that ‘ every negro with 
enlarged glands must be considered, until the contrary is proved, to 
be a case of human trypanosomiasis/ Koch 4 confirmed the value of 
the method in the course of his work, and more recently the British 
investigators in Uganda have done the same. 

As compared with other means of detecting the disease, in the 
absence of definite symptoms, gland puncture is infinitely the best. 
This has been shown so clearly and so often that no stress need be 
laid on it here. While enlargement of the glands does not occur with 
unfailing regularity in every case, the number of these is so 
comparatively small that it does not invalidate the practical utility of 
the method nor the prophylactic measures based on its application. 

In obtaining our results, we have used the classification adopted by 
Dutton and Todd. 5 This schedule is arbitrary, and is one into which 
the personal equation enters to a large degree, but the exact 
determination of the class in which the enlarged glands should be 

placed, e.g., H-’ or ‘ h -/ is perhaps of more academic than 

practical value. Our figures are based on the palpation of the glands 
in the posterior triangle of the neck. 

In the six months, July-December, 1907, some 9,005 natives were 
examined, and of these 1,878 had palpable glands, a percentage of 
20*85, classified as follows : — 


• 

+ 

+ - 

+- 

5 

36 

i 837 


Expressed as percentages of the total number of enlarged glands, 
we have: — 


+ 

i 

+ - +- 

026 

I*gi 97*8l 




83 


or as percentages of population (based on number examined): — 


1 

: + 

-i— 

1 

1 + " “ 

0-0 5 

3 99 

23-99 


The majority (5,000) were from villages closely bordering the 
Luapula, but the results from various other districts visited were much 
the same, so that these figures may be accepted as a fair index of the 
occurrence of enlarged glands throughout the whole country. 

In as many cases as was practicable, gland puncture was performed 
and the juices thus obtained examined microscopically. The results 
were: — 


Class 

Number 

Palpated 

_ 

Number 

Punctured 

Number 

Infected 

% of successful 
punctures 

+ 

5* 

5 

3 

60 

+ - 

36 

30 

0 

O 

+ — 

1837 

297 

o 

O 


From this table it will be seen that in the ‘ h -’ and * -+-* 

groups the result was uniformly negative, while of the five ‘ + ’ cases, 
three harboured trypanosomes. 

These findings point to the conclusions that slight enlargement of 
the glands commonly occurs unassociated with trypanosomiasis, and 
that excessive enlargement, in practice, means * sleeping sickness/ By 
this we mean that such cases should certainly be regarded with 
suspicion, and should be isolated until puncture can be performed by 
a properly qualified person. The number of positive cases we have 
seen is altogether too small-to permit of any dogmatic statement as 
to how great a percentage of * + 9 glands, in this country, harbour the 
parasites. 

As regards Nyassaland, a report sent to us before leaving England 
shows that of 3,467 natives examined in various parts of that 

• One of these, a child 4 years old, had only one gland, measuring 2x1 cm. 
There was no apparent cause for the enlargement. 



8 4 


territory, 26 had ‘ + ’ glands, 409 ‘ H-,’ and 1,406 4 -f-We 

have no knowledge of any of these having been punctured, so that we 
are not in a position to say whether any are infected. Judging, 
however, from our results in Rhodesia, some of the negroes with ‘ + 1 
glands might quite possibly be cases of the disease. While these 
figures are given as the results of the palpation of the glands in the 
posterior triangle of the neck, one of us was informed by one of the 
medical men concerned in collecting the figures that glands in all 
parts of the neck were included. If this is the case with all, the 
figures would need to be corrected, since some of the glands, e.g., the 
submaxillary and suboccipital, are very frequently enlarged, from 
causes other than trypanosome infection. 

It therefore seems clear from the relative frequency of slight 
glandular enlargement, and the uniformly negative findings on 
puncture, that the axiom ‘ glands mean trypanosomes * needs to be 
revised, so far, at least, as Rhodesia and Nyassaland are concerned. It 
might be stated that excessive enlargement of the glands, sufficiently 
marked to bring them into the ‘ + * category, must be regarded as 
meaning trypanosomiasis until the opposite has been proved. The 
practical meaning is that a medical officer is the only person who can 
satisfactorily apply the method of palpation. This will be considered 
in more detail below. 

OCCURRENCE OF CASES OF TRYPANOSOMIASIS 

Three cases of the disease were found in the Luapula division. 
At the time at which they were first seen, all appeared to be in perfect 
health, and presented no other signs of the disease than glandular 
enlargement. In all of them, the glands in the posterior triangle of 
the neck were the only ones which had increased in size. In the 
village of one of the cases GL morsitans was present, but only the 
one case was found. All had a history of having worked in the 
Katanga mines some three to four years previously. When it is 
remembered that human trypanosomiasis is commonly very insidious 
in its onset, and that cases may remain free from symptoms for years, 
there can be little doubt that these cases originated in the district 
mentioned. Contributory evidence that this view is correct is 
afforded by the fact that no indigenous case was found in any of the 



85 


villages along the Luapula, although 67 pal pal is is present and the 
people constantly exposed to their bites. The infection can be 
clearly traced, then, from Kabinda to Lake Kisale, from there south¬ 
wards to some of the Katanga mines* and from these again it has 
been brought into Rhodesia. 

As regards the Northern portions of the country around Mweru 
and Tanganyika, we have at present no personal experience, but 
there is reason to believe that cases also exist there. These would 
not necessarily be introduced from the Katanga although the possi¬ 
bility of this must not be overlooked. Very large numbers of natives 
from these districts have worked in the mines, and since cases have 
occurred amongst those who went from the Luapula division, there is 
just as much reason to expect that cases will be found in all the 
districts from which labour has been drawn. The other point of 
introduction would be from the endemic centres along the higher 
reaches of the Luapula, and from Lake Tanganyika. In 1901, 
imported cases were present at Moliro, 6 in 1902, at Baudoinville, and 
within the last year the disease has been reported as being endemic 
in the vicinity of Vua. As fly exists (67 pal pal is) along the shores 
of Mweru and Tanganyika, as the people have been communicating 
freely, and as there have been numbers of Swahili traders crossing 
from one country to the other with their retinues, cases of the 
disease might reasonably be looked for. There is good reason to 
believe that these are present. 

All these points bear out in a striking manner the correctness of 
Dutton and Todd’s 6 observations on the way in which the disease has 
been carried from an infected to a non-infected region. 

TRANSMISSION OF THE DISEASE 

Wherever sleeping sickness has been found, its distribution has 
been closely related to that of Glossina palpalis. This has been 
accepted as showing more or less conclusively that the disease can 
only be transmitted by this species. While it would be idle to ignore 
the inferences implied by this relationship of the disease and 67 
palpalis , there is little foundation for the belief that this fly only is 

* At present there are a number of cases of human trypanosomiasis in the 
hospital at Ruwe, and until very recently, at all events, at Kambove as well. 

G 



86 


responsible for the spread of the disease, in the light of our present 
knowledge. 

In parts of the Congo Free State visited by the Expedition of 
this School to the Congo, the disease was found to be widely 
disseminated, although Gl. pal pal is was found only very scantily. 
Since, therefore, the numbers of this fly did not appear to account for 
the number of cases, experiments 7 were made with various other 
biting Arthropods—the larva of Auchmcromyia luleola (Congo Floor 
Maggot), Ornithodoros moubata , Simulium and Anophelines -to 
transmit Trypanosoma gambiense. All these resulted negatively. 
The experiments to transmit by means of tsetse flies were also 
unsatisfactory, and although positive results were obtained, the 
number of flies required was so great that it was felt the solution had 
not been reached. In Uganda, 8 the results were similar. Large 
numbers of flies were required for success, and this only followed 
when the interval between the ‘ infecting ’ and the * transmitting * feed 
did not exceed 48 hours. No satisfactory evidence of a developmental 
cycle of the trypanosome in tsetse flies has yet been obtained, and 
it can now be accepted as proved that transmission by mechanical 
means is possible. The importance of this is self-evident. 
Mechanical transmission does not explain satisfactorily the rapid 
extension of the disease which has been observed in many instances, 
nevertheless, whether it is eventually shown that the trypanosomes 
do pass through a cycle in the tsetse fly analogous to that observed 
in the case of so many other parasitic protozoa, and that Glossina 
palpal is is the natural transmitter of the disease, the practical impor¬ 
tance of this accidental, or mechanical, transmission by other species 
of the genus cannot be overlooked. 

In a report sent in to the Administration of North-East Rhodesia 
in 1906, Dr. Noble states that Dr. Polidori, of the Congo service, told 
him that their experience led them to believe that Gl morsitans had 
to be incriminated as well as Gl. pal pa l is. Arguing by analogy 
from the work on cattle trypanosomiasis, this is what would be 
expected. As is well known, Ngana is ordinarily spread by Gl. 
morsitans (and probably pallidipes) yet successful transmission 
experiments have been carried out with a Tabanid; 9 and other 
trypanosomes, naturally transmitted by one species or other of the 
genus Glossina , have been carried from animal to animal by distinct 



«7 


species of the same genus and such entirely different ones as 
Stomoxys 8 and Tabanus .. While arguing by analogy is often a 
fallacious method, some proof that in this particular instance it can 
be accepted is afforded by the recent demonstration in Uganda that 
Trypanosoma gambiense can be transmitted by Gl. fusca. 10 

This question of exactly what species of biting flies, more 
particularly tsetse, are capable of transmitting human trypanosomiasis 
is one of the most important which still remains to be decided in 
connection with this disease. If it can be shown that Glossina 
pal pal is is directly responsible for the spread of the disease and that 
the other species are only accidental carriers, the work of controlling 
the extension will be very much simplified and the cost greatly 
lessened. This is a point which merits attention from all the 
Governments concerned. 

In brief, our knowledge as to the transmission of the disease 
stands thus— 

1. The only known method of transmission is mechanical. 

2. GL pal pal is and Gl. fusca can transmit the disease. 

3. At present all other species of Glossina must be regarded 
with equal suspicion. 


PROPHYLAXIS 

The whole system of prophylaxis is based on the application of 
gland palpation and puncture. Since by this means we are enabled 
to detect the disease in its earliest stages in over 97 per cent, of the 
cases, we are in a position to weed out the infected and isolate them 
before they have become very dangerous. It is manifest that as long 
as the trypanosomes are confined to the glands, as opposed to the 
peripheral blood circulation, the chances of a tsetse fly becoming 
infected are comparatively small. Koch 11 has also stated that the 
tsetse flies he employed only became infected when animals were used 
which had had the disease for a considerable length of time, and in 
whose blood the parasites were scanty. From this it will be seen 
that the value of gland palpation is enhanced so far as prophylactic 
measures are concerned. These measures may be divided into two 
broad sections—1 major, and 2 minor measures. 



88 


1. Major Measures. 

These are—A. Control of native movements. 

B. Segregation of cases. 

C. Removal of villages from dangerous zones. 

2. Minor Measures. 

A. Clearing. 

B. Education of the natives. 

C. Personal prophylaxis. 

D. Destruction of tsetse flies, their larvae and pupae. 

i. Major Measures. 

A. Control of native movements. 

Of their own accord, natives do not move about the country in 
large bands. These are either directly associated with Europeans, 
Swahili, and Arab traders, or indirectly under their control. 
Legislation to control the direction of these movements would 
accordingly do much to prevent the importation of cases from infected 
to non-infected areas. This legislation should make it an offence, 
punishable by suitable fines, for any person having infected natives 
in his employ, taking natives from a non-infected region to an infected 
one and vice versa, or otherwise violating the regulations promulgated 
from time to time with regard to sleeping sickness.* Wherever 
possible it would be well to have all natives who are travelling for any 
distance certified as free from symptoms of the disease by a competent 
person. 

In the case of North-East Rhodesia, the only dangerous 
movements are from Madona to the Katanga mines, and the 
operations of the Swahili trader along the northern border. We 
believe that the cases we have found originated at the Katanga 
mines; in addition there are cases under treatment at Ruwe, and 
until very recently at Kambove as well; and tsetse flies exist along 
the whole of the route from Madona to Kambove. The danger of 
this traffic was pointed out as long ago as February, 1907, and the 
stoppage of labour-recruiting for the mines was then advised. This, 
however, was not done until later in the year, when the receipt of 
reports from Dr. Sheffield Neave, in addition to those previously sent 

* That some such legislation is required is shown by the fact that although 
Madona is now the only place at which the Luapula can officially be crossed, white 
men have on several occasions crossed the river higher up after leaving that place. 



8q 


in by Dr. Massey, made it plain that the disease was steadily gaining 
ground in the southern portions of the Congo Free State. Permission 
has been given to the Tanganyika Concessions, Limited, to recruit 
labour in British territory to transport the loads now lying at Madona 
(6,000 odd), and those actually in transit in the country, to the mines. 
We are of the opinion that this policy is mistaken, and that the 
Tanganyika Concessions should be required to take the loads across 
the Luapula and find the necessary carriers in the Congo Free State. 

On the West, Rhodesia is separated from the Congo Free State 
by a boundary which can be watched with comparative ease, viz., the 
Luapula river. With exception of fords at the Mombatuta Falls, at 
Madona and at the Johnston Falls, the river can only be crossed in 
canoes, and when the river is in flood these fords are impassable. 
The measures suggested, therefore, to protect the river are the 
confiscation of all canoes and the placing of patrols at the fords should 
this be found necessary. To render these measures absolutely 
effective, the co-operation of the Congo Government will have to be 
obtained, for if the villages on that side of the river are allowed to 
retain their canoes, it would nullify to a great degree the benefit 
derived by the confiscation of the canoes on the British side. If the 
mining companies in the Katanga refused to give work to any natives 
of Rhodesia who might get across the river, the general mass of 
natives would soon learn that it was useless to go to the mines, and 
the temptation to leave their villages would thus be removed. In 
addition any uninfected natives of British territory who are in the 
Katanga at present should be returned immediately. 

The operations of the Swahili traders are chiefly confined to the 
northern border. The obvious way of dealing with them is to refuse 
licenses and to require them to leave the country absolutely. 
Regulations to this effect have been passed. The control of a land 
boundary, especially in a country like Africa, cannot be perfectly 
effected, but by the stoppage of organised traffic much can be done. 
Eventually it may be necessary to establish a patrol along the border 
between Mweru and Tanganyika, but the utility of moving all the 
villages from a strip parallel with the boundary might first be 
considered. If practicable, it would do more to stop communication 
than any system of surveillance, however complete. Here 
co-operation with the Congo authorities would be advisable. 



go 


Another region which requires attention is that part of the 
boundary between North-East Rhodesia and the Congo State, 
extending from the Luapula to North-West Rhodesia. One of us 
was informed by Mr. Croad, the Native Commissioner at Serenje, that 
the Congo Free State claimed the territory twenty miles to the East 
of the true boundary, and that in this debated land the Katanga mines 
are recruiting labour. Until the boundary dispute is settled, the 
British and Congo authorities should unite in forbidding any 
recruiting to be done in the country in question. 

Along the frontier between North-West Rhodesia and the Congo 
Free State no natives are now supposed to cross, and as the country 
along this border is very sparsely populated there is not much violation 
of this regulation. The most important point is that all loads going 
to the Katanga mines have to be carried from the frontier by labour 
recruited in that territory. As we have pointed out above, this should 
also be the case in North-East Rhodesia. 

With respect to Nyassaland, no labour is now allowed to proceed 
to the Katanga. While this is so, it must not be forgotten that there 
are there a number of skilled workmen and raw labourers from this 
country, and these will be returning to their homes at future dates. 
It is an open question whether some of those who have already gone 
home have not already carried the infection into the Protectorate, 
just as has occurred in the case of Rhodesia. In a recent number of 
the British Medical Journal, 12 we notice that the Principal Medical 
Officer of Nyassaland does not anticipate the entrance of the disease 
from the North, and even if this be so, a point on which we still have 
some doubts, the possibility of it coming in by the route mentioned 
above must not be ignored. As GL fusca occurs in the Protectorate, 
and as it is capable of transmitting the disease, the danger which may 
result from the introduction of human trypanosomiasis is apparent. 
The chief line of trade from the North end of Nyassa is from Karonga 
to Kasama, in Rhodesia. So far as is known at present there is no 
danger connected with this, though odd cases of the disease, imported 
from the Katanga, may exist in the neighbourhood. Whether there 
is any danger of it coming in from German East Africa, we cannot 
say. Imported cases were present at Udjidji in 1906, and 
GL pal pal is is found along the German shore of Tanganyika to below 
Bismarckburg. 



9i 


B. Segregation of cases. 

This is a most necessary precaution. Cases of trypanosomiasis 
exist in the country and tsetse flies are widely distributed, in one 
instance, at least, being fairly plentiful in the village in which an 
infected man was living. The practical application of gland 
palpation and puncture are the means to be adopted in finding the 
cases. Dutton and Todd 5 pointed out that any European, or even 
intelligent native, could apply the method. While it cannot be 
doubted this is possible, it would be advisable not to rely on the 
chance assistance of either of these. Even with the best of 
intentions, the majority of Europeans would not fully appreciate the 
importance of the issues involved, and would soon tire of practising 
it. A dependence on results thus obtained would only lead to a false 
sense of security. Again, the fact that enlarged glands in this 
country does not necessarily mean trypanosomiasis is another reason 
why the work should be left in the hands of trained medical officers. 
The saving of time where palpation and puncture can be done on the 
spot, and the consequently lessened danger of having possibly 
infected people travelling through the country to the nearest medical 
officer, are facts worthy of consideration. At the present date it 
appears probable that cases are scattered over a wide area of the 
territory; therefore we would suggest that a sufficient number of 
special medical officers be appointed to travel systematically through 
all the villages palpating all the natives and puncturing those in which 
this was indicated. In the event of cases being discovered, they 
should at once be removed to a segregation camp for treatment. 
Before leaving their villages, or, in fact, as soon as the diagnosis had 
been established, they should be given a full dose of atoxyl, 
preferably intravenously. This would drive the trypanosomes from 
the peripheral circulation—Koch 13 states for at least 30 days—and 
would prevent the possibility of infected persons acting as 
disseminators of the disease on their way to the camp. 

It must not be thought that one visit to a village will be sufficient. 
Two of the cases we found only escaped earlier detection by reason 
of the fact that they were absent from their villages when these were 
first visited. 

The districts into which the country is divided for this purpose 
should not be so large as to make it impossible for the medical officer 



92 


to do the work satisfactorily. The villages should be visited at least 
twice a year, though oftener would be much preferable and advisable. 
Too much work would be entailed if one medical office* were 
expected to do all this travelling, and in addition look after a 
segregation camp. To provide two medical men for each sleeping 
sickness district would perhaps be too expensive, though if the 
political divisions (in North-Eastern Rhodesia) are adhered to, this 
number would be required. In North-Eastern Rhodesia the cases 
will be found chiefly along the frontier, and it would be quite possible 
to have one central segregation camp somewhere on the plateau 
which would serve the requirements of the whole Northern portion of 
the country. This would have to be under a resident doctor. 

It must be admitted that this scheme will entail a substantial 
expenditure, but the cost will only be a fraction of the loss which 
would result from a wide-spread extension of sleeping sickness, and 
in addition there is a fair chance of success if put into operation at 
once. Another point is that the expenditure would probably not be 
a permanent drain on the country, as when all the cases have been 
found and isolated, and when the frontiers have been finally closed, 
the staff could be reduced. As more knowledge is obtained of the 
bionomics of tsetse flies, of the fate of trypanosomes ingested by them, 
and of what flies are capable of transmitting the disease, other 
methods of controlling its spread may be found. 

In Nyassaland the disease is unknown at present, though there 
is a possibility of it being in the country already. 

C. Removal of villages from dangerous zones. 

As long as villages are left in places where tsetse flies abound, 
the introduction of a case of trypanosomiasis is dangerous. We quite 
recognise that it will be impossible to carry this measure into effect 
everywhere, but in situations where Gl. pal pal is, at least, exists, it 
should be put into application. In other cases clearing must be 
resorted to. In Rhodesia, this measure would apply to those villages 
lying along the Luapula, Mansa, Luongo and Kalungwisi rivers and 
Lakes Mweru and Tanganyika. Whether it can be done in all these 
situations we cannot say, but with regard to the Luapula, the ‘Mansa 
and the Luongo it is practicable, and in fact is being enforced. In 
most cases these particular villages only settled on the river after the 



93 


advent of British rule had ensured peace ; formerly they were further 
back in the hills, and it is to these former sites that they are being 
moved. This procedure will also lessen the chance of natives crossing 
the river surreptitiously. 

2. Minor Measures. 

A. Clearing. 

In those cases in which the villages cannot be moved, the 
surrounding land must be cleared. The extent to which this must be 
done appears to be variable. Dutton and Todd advise 300 yards, 
while in Uganda, Dr. Hodges 1 states that a break of 50 yards (in 
the case of Gl. pal pal is) is sufficient to banish them from their 
natural haunts. In this connection Madona affords a striking 
example. On either side of the ferry, the whole shore has been 
absolutely cleared for a distance of 200 yards. Beyond this again the 
land is planted with gardens for some distance (about 500 yards to 
the East and 700 to the West). From the river’s edge the clearing 
extends back for 350 yards, and in the middle of this clearing, some 
225 yards from the river, the various residences and offices are placed. 
The river here is over 400 yards in width. It will thus be seen that 
the clearing more than satisfies the most exacting demand that has 
yet been made, but in spite of its extent, specimens of Gl. palpal is 
have been seen and caught on at least half a dozen occasions on the 
verandahs of some of the buildings, and this too when there was no 
possibility of them having been accidentally carried from the bushy 
part of the shore. We would, therefore, consider it more advisable 
to move villages from fly-infested locations than to leave them in 
small clearings. The native is notoriously lazy and careless, and 
even if forced to make clearings, would allow them to grow up again 
unchecked unless continuously supervised. In cases where villages 
have to be left, we would insist on the 300 yard clearing as the very 
smallest that should be allowed. 

B. Education of the natives. 

The relationship existing between tsetse flies and sleeping sickness 
should be explained to the chiefs, and the importance of placing their 
villages in fly-free country. So far as Rhodesia is concerned, most 
of the natives know from practical experience that cattle and sheep 



94 


invariably die when brought into contact with f tuzembe,’ and would 
therefore appreciate the importance of giving these insects a wide 
berth. At present sleeping sickness is unknown to the natives, and 
unless controlled they would not be deterred from pursuing such 
occupations as fishing, by an abstract fear of a disease of which they 
know nothing. The ordinary bush tsetse ( 67 . mor sit arts) is known to 
most of them, as mentioned, but they are not so well acquainted with 
pal pal is. The chiefs might be asked to notify any case of unusual 
illness in their villages and to clear the bush around the places where 
water is drawn. No reliance, however, could be placed on their 
promises to carry out any regulations.* 

C. Personal prophylaxis. 

This is obviously directed to the prevention of the bites of tsetse 
flies. Adequate clothing and the use of some of the means adopted 
in the prophylaxis of malaria would be applicable here, e.g., head nets 
where the fly are very bad. 

D. Destruction of tsetse flies. 

Unfortunately we know of no means of directly destroying them. 
Very little is known of the bionomics of these insects, so that we are 
in the dark as to their most vulnerable point. So far as our 
experience goes, burning the veldt does not make much difference in 
the number of flies. Indirect measures of banishing them, such as 
clearing, have been mentioned above. 

In brief then, the measures we would suggest for adoption in 
Rhodesia are— 

1. The absolute and immediate closure of the Luapula. 

2. The stoppage of transport from Madona to Kambove by 
natives of Rhodesia. 

3. The return of all uninfected natives of Rhodesia, and Nyassa- 
land, from the Katanga mines to their homes. 

4. The various mining companies in the Katanga should be 
requested to refuse work to any native of Rhodesia. 

5. The Rhodesian and Congo Governments should refuse 
permission to recruit labour in disputed territory. 

* For instance, the chiefs along the river have been told that they must not 
take anyone, white or black, across, yet in spite of this such cases have come under 
our own observance. 



95 


6. All the canoes along the Luapula river should be seized. 

7. All Arab and Swahili traders should be expelled from the 
country. 

8. All villages on either side of the Mweru-Tanganyika boundary 
should be moved back, if possible. 

9. Should it be impossible to carry No. 8 into effect, the 
establishment of patrols must be considered. This would, however, 
not be so satisfactory. 

10. Legislation should be passed to deal with any infringements 
of the regulations promulgated with regard to sleeping sickness. 

11. The Government of the Congo Free State should be 
requested to co-operate actively with the Administration of Rhodesia 
in rendering any measures effective which may be adopted from time 
to time for the protection of the frontiers. 

12. That special medical officers be appointed to travel through 
the country to search for cases of the disease. 

13. That a central segregation camp, under a resident doctor, be 
established in a district free from fly. 

14. That all villages be moved, wherever possible, from the 
vicinity of tsetse flies, more particularly Glossina pal pal is. 

15. In cases where removal is impossible, clearings round the 
villages at least 300 yards in width should be insisted on. 

16. The chiefs should be instructed with regard to the disease, 
its relationship to tsetse flies, and the importance of keeping the 
villages in fly-free country. 

Madona, February 1 st , 1908. 


In a letter dated Madona, February 13th, 1908, Dr. Kinghorn 

refers to the official report of Dr. Spillane. He states that the 

report verifies what Dr. Todd prophesied in 1906. He also states that 
GL morsitans , as a possible transmitter of the disease, is ignored in 
drawing up preventive regulations. 

‘ Another case of sleeping sickness has been found in the vicinity 
of Madona. This man, a chief named Matanda, says he has never 
been in the Congo since Europeans came into the country. His 
village is not far away from two others in which cases have been 
found. 




g6 


* He has been a very big man, over six feet in height and well- 
developed. Now he is very much emaciated, can only rise when 
assisted, and walks with difficulty. Has no headache or other 

symptoms. Glands H-, and on puncture show trypanosomes. 

These are also present in the peripheral blood. The case was 
diagnosed by Dr. Storrs, M.O., and I found trypanosomes in the blood.’ 


REFERENCES 

1. Colonial Reports, Uganda, No. 4662, 1907 (proof). 

2. Greig and Gray. British Medical Journal, May 18, 1904. 

3. Dutton and Todd. ‘ Gland puncture in Trypanosomiasis.’ Liverpool School 

of Tropical Medicine, Memoir XVI, 1905 

4. Koch, R. ‘ Ueber die bisherigen Verlauf der Deutschen Expedition zur 

Erforschung der Schlafkrankheit in Ostafrika. 1 Deutsche Med. 
Wochenschrift, No. 51, 1906. 

5. Dutton and Todd. ‘Gland Palpation in Human Trypanosomiasis.’ Liver¬ 

pool School of Tropical Medicine, Memoir XVIII, 1906. 

6. Dutton and Todd. ‘ Distribution .... of Sleeping Sickness in the Congo 

Free State.’ Liverpool School of Tropical Medicine, Memoir XVIII, 1906. 

7. Dutton, Todd and Hanington. ‘ Trypanosome Transmission Experiments.’ 

Annals of Tropical Medicine and Parasitology, Vol. I, No. 2, 1907. 

8. Minchin, Gray and Tulloch. ‘ Glossina falpalis in its relation to 

Tryp. gambiense and other trypanosomes.’ Proc. Roy, Soc., Series B, Vol. 
78, No. B525, p. 242, 1906. 

9. Sergent, Ed. and Et. ‘ Etudes sur les trypanosomiases de Berberie en 

1905.’ Ann. de’Inst. Past., t. XX, p. 665, Aug. 1906. 

10. Colonial Reports, East Africa Protectorate, No. 6490, 1907 (proof) 

11. Koch, R. ‘ Vorlaufige Mittheilung liber die Ergebnisse einer Forschungsreise 

nach Ostafrika.’ Deut. Med. Wochenschrift, Vol. 31, No. 47, 1907. 

12. Report reviewed in British Medical Journal, No. 2444. Nov. 2, 1907. 

13. Koch, R. 1 Bericht iiber die Tatigkeit der deutschen Expedition zur 

Erforschung der Schlafkrankheit bis zum 25 November, 1906.’ Deutsche 
Med. Wochenschrift, Vol. 33, No. 2, 1907 






97 


A REPORT ON TRYPANOSOMIASIS OF 
DOMESTIC STOCK IN NORTH-WESTERN 

RHODESIA 


BY 

R. EUSTACE MONTGOMERY, M.R.C.V.S. 

AND 

ALLAN KINGHORN, M.B. 

THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE EXPEDITION TO THE ZAMBESI, 1907 


(Received for publication io April , 1908) 


SYNOPSIS 

PAGE 


Local Views on the Disease . 97 

Domestic Animals of the Territory . 101 

Trypanosomiasis in Cattle . 102 

Infection by T. dimorphon . i °4 

Experimental . io 7 

Infection by T. vivax . 112 

Experimental . 114 

Infection by T. theilcri ... 117 

Trypanosomiasis in Sheep . 117 

Trypanosomiasis in Native Dogs . 118 

Morphology of the Trypanosomes . 118 

T. dimorfhon . 118 

T. vivax . 122 

Trypanosomes found in Dogs . 124 

Argument . 125 

Transmission of the Cattle Trypanosomes . 128 


The Expedition reached Broken Hill, the present head of the 
Cape to Cairo Railway (approximately 14 0 35' S., 28° 40' E.) on 
June 17th; on the 23rd the examination of some cattle at a farm in 
the neighbourhood revealed the presence of trypanosomes. 

LOCAL VIEWS ON THE DISEASE 

* Fly disease/ 1 fly struck ’ or ‘ fly ’ is well known to the inhabitants 
of the district, and most deaths in cattle are ascribed to it. The 
views held locally are those of stock owners South of the Zambesi. 
The passage of cattle through a known tsetse area is only undertaken 
under compulsion, and wherever possible is effected during the night, 



















9 8 

and it is considered that the bite of more than one fly is usually 
necessary to give the disease. 

Cattle are held to be most susceptible, but cases are recorded of 
animals which do well and breed in villages situated within fly areas. 
There is a fairly general impression that animals born in the fly area 
possess a degree of immunity not enjoyed by others, and this is held 
to account for the herds which some native chiefs are said to own. 
We have seen such a village near the River Kafue, and have taken 
Glossina morsitans within a mile and a half of it. The chief of this 
village regards the freedom of their cattle from the disease as being 
due to care in herding, and admits that if they were taken away from 
the old garden clearings except at night that sickness would be 
expected. Another village was frequently quoted as an instance of 
this inherited immunity, but on arrival there we found that within the 
past two years most of the cattle had died of fly disease, and the rest, 
which had been sold to a European, had also succumbed. We only 
met with one case which would tend to bear out this suggestion of 
local insusceptibility. 

Sixteen head of yearling stock were purchased from a chief near the Kafue in 
1902. These animals had been bred there and Gl. morsitans is found within half a 
mile. For three years they were kept on the purchaser’s farm, within a few 
hundred yatds of tsetse flies, during which time one animal died suddenly, and 
six were sold fat for slaughter. In 1905 the remainder were moved twenty miles 
down the river, and during the rains of 1906-1907 six deaths occurred. Five of 
these had given birth to calves, and these, with the rest of the cows, appeared in 
excellent condition when examined in November, 1907. These cattle had spent 
their lives within a mile of Gl. morsitans which had frequently been caught feeding 
on them. Game is plentiful around. Bushbuck, puku, waterbuck, and hartebeest 
have often been seen grazing a few hundred yards from the homestead. 

The susceptibility of the mule and donkey is disputed. A team 
of the former and some donkeys were kept for transport near Broken 
Hill, and are said to have frequently been in contact with the tsetse, 
but we were not informed of any deaths which could be directly 
attributed to this cause, and an examination of eight of these mules 
did not show trypanosomes. We received the histories of three 
donkeys which are said to have lived in fly areas for from one to three 
years without ill effect One of these was later shown to be 
susceptible to infection by T. dimorphon . A second was taken into 
a badly infested district in August under the belief that it was 
immune. Its blood was examined in November by a layman, who, 
however, had a good deal of experience in blood examinations; and 



99 


it was stated to have trypanosomes. Fifteen days later it appeared 
in good health, and we were unable to verify the finding in the single 
observation made. Dr. Yale Massey showed us films of T. dimorphon 
made from the blood of a donkey which died of the disease at Ruwe 
in the Congo Free State. 

Goats are considered as immune by Europeans and natives alike. 
This view is held to such an extent that they will graze a herd of 
these animals over land intended for occupation in order to drive 
away the tsetse, which is said to be not only harmless to them, but 
also to be repelled by the odoriferous nature of the adult males. We 
were informed of many instances where goats had lived and bred 
within fly areas. From one such area a European purchased forty 
head in February, 1907. They commenced to die immediately at 
the rate of one or two a week, the symptoms being emaciation lasting 
for one or two months. In November three adults (all that remained), 
each of which had a kid, were examined with negative result. 

Native dogs are regarded as immune, and it is stated that natives 
will expose their dogs, a valuable asset to them, without fear. We 
have seen three cases of natural infection in 1 essential kaffirs.* 
English and Colonial dogs are considered susceptible; but if born or 
bred in the fly district their powers of resistance are said to be 
increased. 

No special symptoms are recognised. Gradual and progressive 
emaciation ; periods of manifest depression followed by others in 
which the animal appears brighter ; lacrimation and nasal discharge ; 
these, taken with a history of passage through a fly district, are held 
to be sufficient for diagnosis. Oedema, enlargement of the 
superficial glands or paresis, are not mentioned. On autopsy an 
oedematous condition of the connective tissues and a paleness of the 
muscles are said to be constant; the presence of fluid in the body 
cavities has not always been noted by stock owners. 

Deaths are said to be more frequent just after the break of the 
rains, and it is considered that any undue exposure of an animal to 
water will ‘ bring out * the latent disease; so much is this held 
that we have been informed of cattle being purchased subject to a 
test of pouring a bucket of water over them. Should they not show 
evident signs of sickness within a few days, they are considered as 
free from ‘ fly.* Our observations at Broken Hill, and since, do not 



IOO 


bear out the interpretation of this seasonal prevalence given by 
owners, most of whom think that the deaths will occur soon after the 
commencement of the rains irrespective of the date of infection. The 
more reasonable explanation would be that as waggon transport is 
impossible except in the dry season, the infection takes place between 
May and October; and since the disease is of a rapidly fatal 
character (one to five months), deaths will normally occur about 
November at the time rains are expected to break. Further, it is 
obvious that any debilitating influence, and particularly this sudden 
climatic change, will act as an exciting agent and hasten a fatal 
termination. 

The ordinary method of prevention consists in avoiding tsetse 
country; where this is impossible, animals are driven through at 
night, and are kept on an open grass plain during the day, for owners 
have noticed that the tsetse ( 67 . morsitans) frequents by choice the 
bush. A native method of prophylaxis exists, and it appears to have 
also been used as a curative. Previous to entering a fly area the 
water of the cattle is restricted so as to compel them to drink a bitter 
decoction made from the bark of a tree ( Kangomba ) in which the body 
of one tsetse is placed. At night the animals are kept in a hut and a 
fire made of the young twigs and leaves of a bush (irisafwa\ in the 
smoke of which they remain till morning. The effects of this 
treatment are held to last for three to four days, but daily adoption is 
recommended. One European expressed some faith in it, having 
twice taken an animal into a tsetse zone after fumigation. Another 
put the matter to a more thorough test, and after medication sent 
three cattle into a fly area; they all died within six months. As a 
preventative, fumigation may be to a degree efficacious, for the 
presence of an obnoxious agent would certainly tend to repel the 
attacks of the fly; but we question the correctness of the diagnosis 
in those cases, and they are admittedly few, in which a cure is said to 
have resulted. 

With one exception, cattle owners European and native, have 
incriminated the tsetse to the exclusion of all other biting flies, and 
the views common in South Africa regarding the association of 
Glossina and game are also held, but to a more limited and disputed 
extent. The exception was a native of North-Eastern Rhodesia, 
whose father, one of the greatest chiefs on the Western side of the 




IOI 


territory, had lost some eighty head of cattle within a year. This man 
caught us a Tabanus , closely allied to T. dorsivitta , Walk., and stated 
that his people believed it to be the cause of the disease. While this 
is no proof, the spontaneity of the act indicated these natives’ belief 
in the statement, which coming from such a source is the more 
remarkable, as ordinarily they might be expected to blame the 
recognised enemy, the tsetse. 

DOMESTIC ANIMALS IN THE TERRITORY 

These observations refer to that part of North-Western Rhodesia 
which lies between 28° 40' East Longitude and the River Kafue, and 
more particularly to that area in which one of us travelled. 

Cattle breeding is extensively carried on by the Mashakalumbwe 
people, who occupy the North bank of the Kafue as it runs Eastwards 
to join the Zambesi. Isolated members of the same tribe living to 
the North of the rest are also cattle owners; the other tribes are not 
cattle-men, though a few of the largest chiefs whose villages are 
marked on the map possess a few cows in the kraal. If they own any 
more, these are quartered out with other chiefs resident in fly-free 
districts. It would appear that even these few are discontinuing the 
custom, partly owing to the losses they sustain from the tsetse, and 
perhaps also because with the presence of Europeans the prices of 
cattle have risen, and they will no longer replace losses. As practically 
the whole area North of a line drawn from the South-Western corner 
of the 4 Hook of the Kafue ’ to the South-Eastern boundary of the 
Congo Free State is infested by Glossina morsitans , it is understood 
that cattle-raising to any extent would be impossible, and that its 
absence is not entirely due to the indolence of the native. We are 
informed that in 4 the old days ’ cattle breeding was extensively 
carried on here, and that the spread of the tsetse, the epidemic of 
rinderpest, and the tribal raids that took place before the British 
entered into possession, have brought about its cessation. 

European settlers own one to three spans, each of sixteen to 
eighteen head, for agricultural work ; but the opening up of mines, 
and the lack of other forms of transport, have caused these animals 
to be taken from the farms and utilised for this purpose. Cows are 
scarce, and are now obtained, together with the working bullocks, 
from the Mashakalumbwe on the Kafue, or from the Barotse Valley 

H 



102 


on the Western side of the British sphere of influence, which is said 
to be free from fly and an excellent cattle country. There are a few 
animals of the Zebu type coming from German East Africa, or from 
the Ngoni Country to the East of Rhodesia. 

Goats are kept at most of the larger villages, which are relatively 
few in number. They are of a very inferior strain, small and under¬ 
sized, and would appear to have been in-bred for generations. Those 
in the hands of Europeans are somewhat improved, and the rams have 
been imported from the better stock-raising districts South of the 
Kafue. 

Sheep in North-Western Rhodesia are, so far as we are aware, 
owned mainly by settlers. In North-Eastern Rhodesia, however, 
natives own large flocks, mostly small ill-shapen animals, showing but 
an element of Persian blood. Excepting for the smaller develop¬ 
ment of their tails, which hang straight at the tip, they are not unlike 
the Indian dumbah. 

Equines are limited to a few Europeans. They came from South 
of the Zambesi, and some of the donkeys from German East Africa. 

TRYPANOSOMIASIS IN CATTLE 

Our present observations date from June 23 rd, between which 
date and September 30th, 1907, experimental work on the 

morphology of the parasites and the curative influence of atoxyl and 
mercury was conducted. Since October we have been travelling 
continuously. 

This work was only rendered possible by the kindness of two 
agriculturists whose animals were affected—Messrs. J. F. F. 
Johnson and F. C. Miles—who permitted us to make what use we 
could of their sick spans. Much of the experimental work on 
pathogenicity is due to the liberality of the Administration, The 
British South Africa Company, which, in conjunction with Mr. H. U. 
Moffat, the Superintendent of the Bechuanaland Exploration 
Company, placed a sum of money at our disposal for the purchase of 
the more expensive animals. To the Administrator himself, Mr. R. 
Codrington, and to Mr. Moffat, it is difficult to adequately express 
our indebtedness for the constant and continuous interest they 
manifested in the work. Observations of the effects of the trypano¬ 
somes met with upon the usual laboratory animals was only possible 



through the kindness and generosity of Dr. Arnold Theiler, C.M.G., 
the Veterinary Bacteriologist to the Transvaal Government at 
Pretoria, and of Dr. Robertson in charge of the laboratory of the 
Medical Officer of Health at Cape Town. Finally, our thanks are due 
to every European in the neighbourhood for the constant courtesy 
they extended, and for the many demands made on their time in 
supplying us with the result of their experiences; to the 
agriculturists who, without exception, placed their animals at our 
disposal for examination; and to the various Government Officials 
who did all in their power to forward the research. 

We established a temporary camp and laboratory near Broken 
Hill, and during our stay there, 36 cases of trypanosomiasis in cattle 
were detected, of which 29 were under continuous observation. This 
work was discontinued on September 30th, in order to prosecute our 
enquiries further North in accordance with the scheme drawn up 
before leaving England. 

The history of many of the 36 cases was uncertain, save that at 
some period within the past six months they had been in a fly district. 
One herd, however, possessed peculiar interest as indicating in the 
most convincing circumstantial manner that biting flies other than 
Glossina can transmit infection. This will be discussed under 
‘ Transmission/ 

We have already noted that with the opening up of mines, cattle 
previously and primarily intended for agricultural work have been 
utilised for transport, and on primd facie grounds this exposes them 
to greater risks of infection by bringing them into contact with the 
tsetse’s haunts. At the present time one of the four ' roads ’ from 
Broken Hill must be used for waggon transport. The two which run 
North enter a fly district about fourteen miles out; the third and 
most frequently used one passes through a patch in which tsetse are 
generally seen, about eight miles South, but being a narrow zone, 
night marching may avoid attack. The fourth runs for 112 miles 
South-West to the copper properties there; only within this last year 
have tsetse been found encroaching at about the 100th mile. This 
road joins that coming North from Kalomo, which was used prior to 
the railway. From the neighbourhood of the junction a road runs 
North to the copper mines, and is in fly-belts most of the way, and a 
new one has just been cut in a South-Easterly direction to join the 



railway some 30 miles North of the Kafue. This runs most of the 
way on a watershed; Gl. morsitans has not yet been seen, and the 
nature of the country is not indicative of its presence, but on all the 
other roads the danger of infection is constant, and when the 
Northern ones are used it is recognised that all cattle must be 
sacrificed. 

In the first herd examined the trypanosomes seen in fresh cover- 
glass preparations appeared to be of two varieties. One, the 
prevailing type also found in other herds, was seen to possess the 
morphological features of T. dimorphon , Dutton and Todd. The 
second was seen in four of these animals, and later in a fifth coming 
from another herd, and in two cattle which had been exposed to 
tsetse-flies experimentally. This, by reason of its extraordinary 
rapidity of motion in cover-glass preparation, is regarded as allied to 
T. vivax, Ziemann. 

INFECTION WITH T. DIMORPHON 

(1) The natural disease in Cattle.—There is nothing in the 
clinical picture of this disease to differentiate it in any way from other 
forms of trypanosomiasis. At some period there is an appearance 
of emaciation and dulness, the coat harsh and hide-bound, head 
drooping, eyes dull and watery, but petechiae on the conjunctival 
membranes were very rarely noted, and occasionally there is a nasal 
discharge. Weakness or paresis of the hind limbs is not common, 
and oedema was not seen. Enlargement of the lymphatic glands, 
notably the prescapular and precrural, is constant, but is of little 
diagnostic importance owing to its prevalence in apparently healthy 
animals in which trypanosomes could not be found on blood 
examination or gland puncture. 

All animals under observation were placed at night into the 
ordinary cattle kraal of the country—an open enclosure fenced to a 
height of seven or eight feet to avoid the attacks of wild beasts—and 
were allowed out to graze all day. Temperatures were taken and 
blood examinations commenced between 8 and 11 a.m., and the 
temperature taken in the evening between 4-30 p.m. and sunset. For 
these operations the animals were brought into a kraal-like enclosure 
leading to a 1 crush,’ into which each was in turn driven. After the 



first day the cattle accommodated themselves to this method, which 
cannot be held as interfering with the proper thermometric 
registration. Under these conditions the normal temperature of 
healthy cattle was found to be approximately ioo° to 102° F., two 
degrees representing the normal diurnal variation. 

During the course of the disease the temperature was almost 
constantly elevated; to a slight degree it was paroxysmal, but, as the 
accompanying charts show, this was not a marked feature. 

Parasites could usually be found on direct examination of 
peripheral blood. For graphic representation a similar system to 
that adopted by Lingard is used, but the number of trypanosomes 
present is so much less (12 to a field (Zeiss Oc IV, Obj. D) was the 
greatest number seen) that lower values are accorded to each mark. 
When numerous it was customary to count forty fields and take the 
average; but when scanty sixty to a hundred, according to 
indications. No animal is marked ‘ absent * unless this latter number 
was counted; and. in necessary cases, as in the treatment 
experiments, a f-inch square cover-glass was searched before placing 
a minus sign. 

There is no close relationship between the temperature and the 
number of parasites seen; sometimes a temperature of 104° or 
105 0 F. was unaccompanied by trypanosomes, or only one to twenty 
fields; and again, a temperature of 101 0 has been seen with four 
organisms to a field. Two cases were observed in which organisms 
were not seen for two or three weeks, and in one (No. XIII) which 
was diagnosed on July 18th, trypanosomes were only seen on 
August 10th (one to a cover-glass during the ten weeks the animal 
remained under observation). These might be considered as chronic 
or latent infections, though both died, apparently within the usual 
time of the disease. 

Gland puncture of the prescapular lymphatics was tried as an 
additional aid to diagnosis, though its general employment is not so 
generally necessary owing to the fairly constant presence of trypano¬ 
somes in the peripheral blood. The method is essentially that 
described by Dutton and Todd for the diagnosis of human trypano¬ 
somiasis, all specimens being sealed with vaseline and examined 
immediately. It will be seen from the figures below that it is of less 
value than blood examination as a means of diagnosis, but its 



io6 


employment is advisable before any suspected animal be considered 
free from the disease. 

Animals in which Animals not 

tryps. were found showing tryps. 
in peripheral blood peripherally 

Number of glands punctured ... 26 ... 33 

Tryps. seen in gland juice ... 19 ... 1 

From a few observations made in India on camels suffering from 
Surra we considered that this method was a valuable aid to diagnosis, 
and in this respect we agree with the views of Dutton and Todd, 4 who 
used it in the detection of trypanosomiasis in cattle of the Congo; 
but in Rhodesia our results so far lead us to consider this method as 
of little diagnostic importance. 

The duration of the disease cannot be definitely asserted, but from 
the views held by local stock-owners and our own observations on 
animals whose histories are fairly complete, and on experimental 
cattle, from one to five months would appear to represent the normal. 
In fourteen of our cases the time which elapsed between diagnosis 
and death averaged thirty days, and many of these animals were in 
excellent condition and could not be suspected clinically. It seems 
probable that in a few cattle a chronic form is established; of this 
we have no direct evidence, but the histories of some stock examined, 
and the occurrence of two herds in which a large percentage were 
clinically cases of trypanosomiasis, but which did not show organisms, 
would indicate the possibility. In one animal of this nature 
(No. XXVII) every sign of the disease from a clinical standpoint was 
shown, but trypanosomes only appeared five days previous to death, 
after an absence of 15 days. 

The lesions observable post-mortem are those of an emaciating 
disease—paleness of the visible mucous membranes, flaccidity and 
pallor of the muscles. The amount of fluid in the body cavities 
varied. In some cases it was practically nil , whilst in one animal 
approximately 1750 c.c. were obtained from the peritoneal cavity. 
Petechiae are present on most of the serous surfaces, notably that of 
the spleen. The blood clots to a considerable degree, and the white 
clots common in equines with surra were not seen. All lymphatic 
glands are enlarged, particularly the precrural, and some of the 
mesenteric, which are also frequently haemorrhagic. Enlargement 



lo; 


of the spleen is inconstant, but it is usually friable with prominent 
malpighian bodies, and the capsule studded with petechiae. In only 
one case. No. XLI, showing marked nervous symptoms, were any 
gross changes seen in the spinal cord. 

Trypanosomes were not constantly seen in the blood at death, nor 
were films, made direct from the lymphatic glands, invariably positive. 
Those, however, made from the haemorrhagic lymphatics of the 
mesentery possessed greater diagnostic value than the others, and 
owing to the danger of a negative diagnosis being made on a blood 
film sent in for opinion, we would recommend the forwarding of one 
made from a haemorrhagic gland in addition, though it must be 
understood that it is necessary for this to be made as quickly as 
possible after death in order to avoid undue phagocytosis. 

These cattle, compared with those of India, are singularly free 
from organic changes in any organ, and from other animal parasites. 
In most (16 in 22) there were a few Filaria papillosa , and in many 
(10 in 22) Paramphistomuni cotiicum was seen. Distomum 
hepaticum was not encountered despite the prevalent idea that 
1 fluke * occurs here. No filarial embryos were seen in the blood, and 
Piroplasma bovis was only seen on one occasion in two animals. 

(2) The experimental disease.—The strain of T. dimorphon 
mainly used in these experiments was derived from a naturally 
infected cow, Case No. XXV. This animal was selected as 
representative of the disease; as she had been confined to the 
homestead and had not travelled on the surrounding roads, the 
danger of a mixed infection being thereby reduced. Though 
apparently in perfect health, she had, owing to her contact with the 
sick, been subjected to four examinations between June 26th and 
July 26th, when she was brought in for daily observation. Trypano¬ 
somes were first seen on July 29th, and it is believed that she 
contracted the disease on the farm. The same strain passed through 
Case No. XLI I served for most of the other inoculations. 

1. CATTLE. Both the animals inoculated with virulent blood 
were later utilised in the experimental work on treatment, which was 
regarded as of more direct importance. Organisms appeared on the 
seventh and eighth day following inoculation, and remained constant 
until the exhibition of atoxyl, in one case, No. XLII, for 24 days. 
Death in the case of No. XLI was largely due to atoxyl-intoxication. 



io8 

The other animal is stated to have died suddenly 63 days after 
treatment was commenced, and 87 days after inoculation. 

Case No. XLI.—August 20th. Inoculated subcutaneously with 2*0 c.c. 
citrated blood of XXV. Trypanosomes first seen August 27th, one to a cover-glass. 
Temperature remained but slightly elevated till the morning of the 28th, when it 
rose to 104° F., and was 105° the same evening. Organisms increased in numbers 
up to the 30th (three to a field) when 5*0 grm. atoxyl were administered and the 
normal disease no longer continued. 

Case No. XLII.—August 20th. Inoculated as above. The temperature on the 
27th evening was 104° F., and the following morning trypanosomes were seen, 1 in 
2 fields. These remained constantly present and in considerable numbers for 24 
days. During this period the temperature was between 102° a.m. and 106*2° p.m. ; 
the animal rapidly lost condition, and showed evident clinical manifestations of 
the disease, and we consider that if treatment had not been attempted death would 
have occurred within two or three weeks at the outside, (vide chart i.) 

Two other cattle, Cases Nos. XLIV and XLV, which were infected by 
exposure to the bites of Glossina morsitans are described later under 4 Transmission.’ 

2. Donkey. 

Case No. LV.—This animal is said to have lived in fly districts for upwards 
of three years and was considered immune. September 3rd, inoculated 
subcutaneously with 2*0 c.c. citrated blood of XLII. During the four weeks it 
remained under observation there was no change from normal in its temperature 
and daily direct examination of blood and three centrifugal examinations did not 
reveal organisms. 

On November 20th it was again examined and T. dimorfhon found, three to a 
cover-glass. It did not at this time show any clinical indications of the disease. 
Since that date we have not received any report as to its condition. 

3. SHEEP. The disease in these animals and goats is of 
considerable interest, for in the quite characteristic temperature chart 
we have evidence for distinguishing this trypanosome from that 
which we hold resembles T. vivax , Ziemann. Three sheep were 
inoculated ; in two healthy animals the incubation was seven and 
eight days, in a third, which had passed through a previous infection 
by the other trypanosome, it was eleven days. 

In both healthy animals the temperature assumed at first the type 
produced by the benign tertian form of malarial parasite in man, later 
giving place to the quartan form. The regularity of the fever is 
striking, as also is the fact that each exacerbation was accompanied 
by an influx of trypanosomes into the peripheral circulation. In order 
to demonstrate this periodicity of trypanosomes and temperature, 
these animals were examined every three hours for a period of 
56 hours, for as we are unaware of any other form of trypanosomiasis 
in which this feature is so pronounced, an effort was made to 
ascertain, if possible, any developmental cycle undergone by the 



ioq 

parasite in the sheep. The time at our disposal, however, was not 
sufficient for a careful study of the question, and our observations 
were without definite results. 

No special symptoms were noted. During the course of the 
disease a rapid emaciation took place, but all the animals were alive 
when the work was concluded at the end of September. Each access 
of fever was manifested clinically by the weakness, depression and 
excessive lachrymation shown by the animal. Oedema was not seen 
in either these animals or goats. 

Parasites were never numerous: on one occasion only were as 
many as six to a field seen. The number, however, depends upon 
the period in the onset or decline of the paroxysm at which the 
routine examination happened to be made. 

Case No. XXXVI.—An aged ram, fat-tailed variety, purchased locally, but 
probably imported from the South. August 7th, inoculated subcutaneously with 
5*0 c.c. citrated blood of XXV. The temperature became irregular on the 5th, and 
organisms were first seen on the eighth day, August 15th. For the first fourteen days 
the paroxysms were tertian in type, afterwards becoming less regular and approach¬ 
ing the quartan type. Emaciation was rapid, but the animal was still alive on 
November 20th, and organisms were present, (vide chart iii.) 

Case No. LVI.—Two-year-old female. September 3rd, inoculated sub¬ 
cutaneously with i*o c.c. citrated blood of XLII. Organisms first seen on the 7th 
day, September 10th, and the initial thermal paroxysm occurred during the night 
of the nth. On the evenings of September 14th, 16th, 18th, 20th, 22nd, and 24th, the 
temperature exceeded 106° F., and on the 28th it reached 107°. During the twenty- 
one days of observed disease the type was essentially the same as in No. XXXVI, 
the chart of which is reproduced. 

Case No. XXXIII.—Male, aged one year. Between August 19th and September 
2nd this animal had shown what we regard as T . vivax with which it had been 
inoculated. After this latter date the temperature remained about normal and 
trypanosomes were not seen. 

September 17th, received subcutaneously 0*5 c.c. heart blood of guinea-pig, 
Case XLVI, dead with T. dimorfhon. The temperature was faily constant until 
the 27th, when it commenced to rise. On the 28th, the nth day, T. dimorfhon was 
seen, the first paroxysm occurring the same day, the second (107*2°) on the evening 
of the 30th, the last day of observation, (vide chart ix.) 

4. GOATS. These animals, as already stated, are regarded as 
immune. Three readily took infection with T. dimorphon after an 
incubative period of seven to twelve days. The disease is of 
essentially the same type as that in sheep, viz., a tertian and quartan 
febrile reaction and a concomitant influx of organisms into the 
peripheral circulation. No special symptoms were observed. 

Case No. XXXV.—Male, aged one year. August 7th. Inoculated under the 
skin with 5*0 c.c. citrated blood of XXV. The temperature became irregular until 
organisms appeared on the seventh day, August 14th. During the seven weeks 



of ia v^hKli yijoptat Aas *>b^toved 4 paroxysm* occurred ynlti lb<f 

regiiWuX' i«! those in Sheep, only the thermal teaethm brimg more prOQht*it?eti— 
108-^ fc\ >i»n ‘LttiK -t>u'/ksk>ji Kaiacialifin was rapid,, the hkd iu tim 

uf om and a halt months that >lap?^d pre-vtous to 'S*r*$mpix soffc ' At* 
evact d^£'-yy^?shtf(eh>ai n if bly no tvim»pe&<v had been t m the farm. {vtd«£ bHati 'ivvjy 

<^>tv N*o rvu-4h?)f, a*?d p.vo y-ais.. Septfinhev 3rd ^u-ri-hMi' 

*ei be busty with* re e£fm?e 4 blood of XUf C*f£afM>a>$ wcrTt.fost 

on (heoutfit H xU>\ Qzpi&iftbtil mb. and p^rovyMyis htriirred oti the -*«ih t t ’hb/f&th, 
aoitr/^th, sA*de v/i " ' 

Yoiih^ mam, -In6e*nXatedr Qn -Augt»*t ^U> with bldAd jiih Coal, 
No.. XI, containing T. vivax\ t'»y}i:jm»Miim\< were never soea, and on September 
17th ft **n ::wed n*s hrart bfood of gniuea-pty, No X! .'VV ; itywol • hi - 

■jji dimafphen. 

On September .29th, the- twelfth day, T dintfirfh^n W4* 3*6ity and again- it*: 
following' day the last of.observation. On November aoth partiMie^ were still 
for^e-nt. and lli^ anVnini Had lost Miiub gf »i^ «:nndjtion. 

5. Dam. Three 5h‘UviN dogs wort- inoculated with this strain, 
of /'. rf;K-h<> t>h->n. iitsii i'n-spoe iiu’ suggested immunity of die race, al! 
took the jliidl we shall later note that naturally 

inflected' native ifogis were rngt with, in which, however, the parasite 
did noi >ho-v the• o.iotp)idk*gm».l • characteristics of T dmwrfhon. 

iti all ; nttCCs i lie disease Avan acute; death taking place within 1 wv>. 
week* v4 the, iippealanck oforganisin', ih aii adult, and withm ten days 
hi young animals of three months old. 

EmacBifkW.is rapid; the cokt becomes harsh, eyes ,lachrymose, 

and hi the adidh opacity o! Ilfh cornea wu$ noted.«lirec days he fore 
death. Superficial lymphatic glands, notably the . piescapular, 
become; enlaced .and;soft, but not pamfui, a feature, however, noted 
in cfei-fernt apparently healthy, dogs. Only doe ghihd pwieitjje wits 
made rVufke ease of ap adult,, -vlren i ijtgfitusms -.vefp not seen m fbe 
peripheral blood ; hot its employment did not reveal-trypanosiMnese 
Nervous symptoms were noted in one, young dog, No F. j V, \vhidv 
•■•.howird parHal iticoordinal ion of ilie hind limbs pp the day- preceding 
■tenth. The incubation period was sixteen days m adult, and seven 
' .md ekseti respeou vcly- ni.young animafe: The:,terfipemture curves 
of t hese bit l ev are most irregular , hardly ever elevated, hut showing 
a -tendency to. liecduie sub normal. In the older dog tire type is that 
of other trypano?.oim,'i:-w'S. -diotyrng iciose resemhlance to many charts 
-■f cahtrres infected .'.vith 7 . Organisms increased progressively 

tip to death. In the adult the temperature and trypanosome curves 
-.'.synchronised- 

Cast? Sck X X X i V.--- ^ bHrh, r<gm:l filiokit two yeam, r\H\ 

!inor'iifgfed With o r, c*trattfd blood of XXV 







Ill 


remained normal until the morning of the 21st when it commenced to rise, 
continuing to do so gradually till the morning of the 26th when it was 105° F. 
That evening it was 102°, and it kept normal for three days to rise again for the 
second paroxysm on the 30th. Trypanosomes appeared on the third day after the 
temperature rose, August 23rd, one to a cover-glass, and increased progressively 
to the 25th, one to six fields. On the 26th, 27th, 28th, they were not seen ; they 
reappeared on the 29th, and were twelve to a field on September 1st. They 
gradually fell in number towards death, which took place on the night of the 4th. 
(vide chart vi.) 

Case No. XLIII.—A ‘kaffir' pup, aged three months. August 20th, 
inoculated 0*5 c.c. citrated blood of XXV. The temperature was very irregular 
throughout, varying from 96*4° to 103-8°. Trypanosomes appeared on the seventh 
day, and remained constantly present till death on the fifteenth day. During the 
last three days they were very numerous, ten to a field, and the temperature was 
usually subnormal. 

Case No. LIV.—Brother to the last. September 8th, inoculated 
subcutaneously with i*o c.c. citrated blood of XLII. The temperature showed 
considerable daily variation. Organisms appeared on the eleventh day, September 
14th, one to five fields, and remained present in slightly increasing numbers, till 
death on the eighteenth day after inoculation, September 21st. 

Post-mortem appearances .—In all three animals the most noticeable feature was 
a considerable enlargement of the spleen, which in the adult dog (weight about 
30 lbs.) measured 33 cm. in length, 10 cm. in width, and 4*5 cm. in thickness; dark 
in colour, sofe and friable and edges rounded. Lymphatic glands of the mesentery 
enlarged congested and haemorrhagic. Petechiae studded the serous membranes; 
the liver pale and friable. The pericardial sac contained from 20 to 75 c.c., and 
the peritoneal cavity of Case LIV 250 c.c. ; the amount in the thoracic cavity was 
not greatly increased. 

6. Rabbit. Only one animal was inoculated. The disease in 
this was of chronic nature, and organisms were rarely seen in an 
ordinary cover-glass examination. 

Case No. XXXVII.—August 7th, inoculated intraperitoneally with i*o c.c. 
citrated blood of XXV. During the four weeks during which it was taken, the 
temperature showed no gross variations. Organisms appeared on the tenth day, 
one to a cover-glass, and on the following day five to a cover-glass were seen. 
They were again detected on the fourteenth, fifteenth and thirty-second day after 
inoculation 5 from this time they were not seen on the daily examination up to 
September 30th, nor on those made about every fifth day since. During the first 
month of disease the rabbit lost condition ; hair began to come out at the base of the 
ears, around the eyes, and on the rump, and sores formed on both tarsi. No 
oedema was noted, nor did any opacity of the cornea or signs of paralysis occur. 
During the next three months the rabbit appeared to improve in condition, and the 
tarsal ulcers dried up. Towards the last fortnight a slightly purulent discharge 
collected on the nostrils and around the eyes. Death took place in a very emaciated 
state on January 21st, 1908, 168 days after inoculation. 

Post-mortem .—The spleen was considerably enlarged and rounded, but firm in 
consistency, the mesenteric lymphatics swollen but pale. Trypanosomes could not 
be found on direct examination; no sub-inoculations were made. 

■j. GuiNEA-PiGS. Five guinea-pigs were inoculated with this 
trypanosome. In all, the disease was of a rapidly fatal character, the 



112 


average duration after appearance of organisms being only twelve 
days. The incubation period in three cases was nine days; in the 
other two, sixteen and nineteen respectively. Organisms were 
constantly present after detection, and were numerous up to death, 
which occurred ten to fourteen days later. 

The temperature in the one animal taken varied little from normal 
until the trypanosomes appeared, when it rose and continued slightly 
elevated to death. The loss in condition was rapid, and the visible 
mucous membranes became very pale. Conjunctival discharge was 
not a constant feature, and in no case was any indication of paralysis 
noted. 

On post-mortem, the spleen was much swollen, measuring from 
5*5 to 7 cm. by 3*5 to 4 cm., congested, rounded, soft and friable, with 
an average weight of 14 grammes. In one case rupture of the capsule 
had accelerated' death. The lymphatic glands were enlarged, 
oedematous and congested. 

9. WHITE Rats. Five of these animals were inoculated. The 
period of incubation varied between six and ten days, and trypano¬ 
somes remained constantly present till death, which occurred in from 
18 to 29 days after inoculation. 

On autopsy, the spleen was enlarged, congested, soft and rounded, 
and the minute mesenteric lymphatic glands were distinct and 
haemorrhagic. 


INFECTION WITH T. VIVAX 

At the examination of the first herd on June 23rd, we noticed in 
one bull the presence of an organism, whose extraordinarily rapid 
passage from edge to edge of the cover-glass, and the transient 
corpuscular displacement produced, caused us to consider it as a 
spirochaete. When our camp was established this animal was 
brought under daily observation, and stained films, and later, cover- 
glass preparations some hours old, showed the organism to be a 
trypanosome. In the same herd one other animal was seen to be 
similarly affected, and two in which T. dimorphon had been found 
showed an occasional parasite whose rapidity of motion approximated 
to that of the bull. Three days before leaving Broken Hill we 
examined 14 cattle not previously inspected, one of which showed the 
same organism. 



(i) Natural infection.—It would appear that stock-owners do not 
recognise more than one form of ‘ fly disease,’ and that the animals 
infected by this trypanosome are considered as cases of ordinary fly 
disease. Of these five animals, that first seen was kept under 
observation for ten days at the original camp, and was later 
purchased and brought to that at which our experimental work was 
* conducted. 

Both animals which showed a mixed infection died within three 
weeks of diagnosis. The one detected just before leaving Broken 
Hill was alive, but considerably thinner six weeks later ; whilst the 
fifth was reported to have died in August, seven weeks after parasites 
were seen, at which time it was in good working condition. 

Case No. VIII.—Bull, aged about five years, (vide chart vii.) No very 
reliable history of this case is available, but it appears that this animal, with the 
rest ol the herd, had been grazed away during the Yainy season of 1906-1907, and 
had passed through fly areas on the way home in February, and that since that date 
it had gradually lost condition. 

On examination, it was emaciated and hide-bound, signs of excessive 
lachrymation were present and the conjunctivae showed a few small petechiae, a 
condition seldom noted in animals infected with T. dimorfhon. Prescapular and 
precrural glands enlarged; abdomen tucked-up ; no oedema, but the bull presented 
all signs of trypanosomiasis, from which it was suspected of suffering by the 
owner. The case was under observation for 26 days between July 10th and August 
15th. 

A noticeable feature in the temperature is the great daily variation, as much 
as 5*4° F. being met with. By joining up the evening or morning registrations the 
‘ curve * produced would not be great; there is no suggestion of a paroxysm, and 
the mean of morning and evening temperatures would not vary much from normal. 

Trypanosomes are always scanty, from one to ten to a cover-glass was the 
usual number, only exceeded on four occasions, when approximately one in ten 
fields was seen. The great rapidity of the organism rendered any degree of 
accuracy in counting impossible, for their presence was generally only ascertainable 
by the slight displacement of corpuscles which accompanied their hurried passage 
across the field, a passage it was impossible to follow by any ordinary movement 
of the slide. With such an organism it is very probable that the one or two 
organisms present in a preparation would not come within the field of vision. On 
two occasions when they were not seen, centrifugalisation showed them to be 
present, and in one case a trypanosome was found in a thick-stained film. We 
would consequently suggest that the trypanosomes were almost constantly present 
in the peripheral blood and that with this parasite there is no great paroxysmal 
increase, and their detection is to a more considerable extent a matter of chance 
than is usual in the other forms of Trypanosomiasis. 

During the 25 days of observation, parasites were seen on 16 occasions, eleven 
were marked + , one -f + , and four + -f+, three of these latter occurring in the 
last week of life, and the other after the animal had been driven 22 miles in two 
days to the camp. Two days previous to death extreme weakness was manifest, the 
patient falling down several times and showing great difficulty in rising. 

Post-mortem commenced 15 minutes after death. Rigor mortis distinct, 
mucous membranes pale, and a few petechiae on one conjunctiva. Skin closely 



1 x 4 


adherent, subcutaneous tissues firm and dry. Muscles pale but firm. Thorax. 
Only a few c.c. of a straw coloured fluid present. Lungs normal, save for slight 
emphysema in both apices. Heart normal in size; no fat in auricular-ventricular 
furrow; muscular tissue pale and streaked with fatty degeneration. Blood dark 
with half-formed clots in each chamber. Abdomen. Approximately 50 c.c. faintly 
blood-tinged fluid. Serous membranes show a few petechial haemorrhages on both 
parietal and visceral surfaces. Fat practically absent. Liver pale, slightly fatty 
and friable. Gall-bladder distended, bile of normal colour and viscid. Spleen , not 
enlarged. Capsule firm, somewhat fibrous and studded with small haemorrhages. 
Malpighian bodies pale, the rest of the splenic pulp pale, lying within the unduly 
defined trabeculae. Kidneys pale, capsule strips readily. Other organs normal. 
F. papillosa and P. coni cum present. 

(2) The experimental disease.—The organism for this work came 
from the bull No. VIII, whose history has just been given, and from 
several animals inoculated from it. 

1 . Cattle. 

Case No. IX.—Calf aged nine months, (chart viii.) July nth, 1907, 
inoculated subcutaneously with io*o c.c. blood direct from VIII. This animal was 
under observation for ten days after inoculation, during which time the temperature 
remained about normal and parasites were not seen. Daily examination was 
recommenced on August 2nd, when it arrived at the new camp. 

Trypanosomes were not seen on arrival, but appeared the following day, and 
as with case VIII were intermittently present until death, which occurred 61 days 
after inoculation. During the 38 days following detection of organisms, these were 
marked + on seventeen, on four, and -f + + on four, and on thirteen occasions 
they were not seen. Two days before death, when the animal was very weak, they 
were extremely numerous, almost swarming across the field, but still retaining 
their great motility. 

The temperature chart is of the same nature as that of the bull; daily 
variations of 3 0 to 5 0 F., without paroxismal tendencies except during the first 
week after arrival at the new camp, which might possibly be that accompanying the 
initial Influx of organisms. 

The loss in condition was gradual, but not excessive, and to the end the animal 
presented a bright appearance despite a progressive weakness, marked during the 
last few days. There was but little lachrymation, but petechiae on the 
conjunctival membranes were observed on several occasions. 

Prescapular and precrural glands were enlarged at the time of inoculation, and 
did not show any perceptible increase in size. 

The animal was accidentally killed by a leopard. 

2. Donkey. One animal only was inoculated, and during the 
twenty-four days of observation organisms were not seen. 

Case No. LVIII.—September 6th, inoculated subcutaneously with 2-0 c.c. 
citrated blood of IX. The temperature rose to 103*2° on the evening of the same 
day and the animal showed slight abdominal distension and pain. It was again 
102° on the evening of the sixth day, and the donkey was somewhat dull with a 
watery discharge from the eyes. It appeared well the following day, and showed 
no further symptoms up to September 30th. During this time no trypanosomes 
were seen, nor did they appear present on November 20th, when the animal still 
retained a healthy look. 



“5 


3. SHEEP. Four sheep were inoculated with this trypanosome, 
and in three the organism reappeared. Only one, a small weakly 
animal, died during the period they were under observation ; and from 
an examination of the temperature charts and of the animals 
themselves, it would seem that recovery may occur naturally, though, 
of course, we were unable to prove this in the time. The inoculation 
period in two cases was seven days. Trypanosomes were always 
scanty, rarely exceeding four to a cover-glass, their activity being as 
great as in cattle. This fact may account for the irregularity of their 
detection and the apparent lack of relationship between their 
occurrence and a thermal rise. The picture presented by the 
temperature chart differs absolutely from that following inoculation 
with T. dimorpkon. The temperature rose between the fourth and 
seventh days, and fell again a few days later, and remained irregular 
for two or three weeks. There was no periodic exacerbation of a 
tertian or quartan type. 

Case No. X.—A small ‘weedy’ animal of five months old. July nth, 
inoculated subcutaneously with 5*0 c.c. blood direct from Case No. VIII. The 
temperature rose on the fourth day and continued elevated for the ten days it was 
under observation at the first camp. Mr. Johnson, on whose farm the second camp 
was established, kindly visited the animal nine days later and made films in which 
trypanosomes were detected. It was then weak, and the native in charge said it 
had been sick for three days. It was carried in, 22 miles, to our new camp and 
died almost immediately on arrival. 

Post-mortem .—Trypanosomes were not found in the blood or gland juice. 
Visible mucous membranes pale, those of the conjunctivae showing a few 
petechiae. The subcutaneous lymphatic glands were enlarged, those of the 
mesentery being also haemorrhagic. There was no excess of fluid in the body 
cavities. Small petechiae studded the pleurae and pericardium. The spleen was 
not enlarged, its capsule was firm and showed a few petechiae. The liver and 
kidneys appeared normal. Neither intestinal parasites, nor D. hefaticum were 
found to account for the debilitated state at the time of inoculation. Two specimens 
of Cysticersus tenuicollis were present in the mesentery. 

Case No. XXXIII.—A healthy sheep aged one year. August 7th, inoculated 
subcutaneously with i*o c.c. citrated blood of IX. The temperature rose to 105*2° 
on the morning of the sixth day and organisms were seen the next morning. During 
the succeeding six weeks the temperature was four times above 105°, and 
trypanosomes were seen on eleven occasions. After September 2nd the temperature 
remained about normal and parasites were not observed. On September 17th it 
was inoculated with T . dimorfhon. (vide antea chart ix.) 

Case No. XXXIX.—A healthy sheep aged one year. August 9th, inoculated 
under the skin with i*o c.c. blood direct from goat, Case XI. Trypanosomes 
appeared on the seventh day, and for the next three weeks were fairly constantly 
present, but during the last three and a half weeks of observation they were only 
seen seven times. The temperature rose on the sixth day, and during the ensuing 
forty days of disease was similar to that of bovine piroplasmosis in India. 



iiG 


Case No. L1X.—A companion sheep to the last. September 5th, inoculated 
subcutaneously with 1*5 c.c. citrated blood from IX (parasites present). The 
temperature remained normal and trypanosomes were not seen during the twenty- 
four days it continued under observation. 

4. GOATS. Three goats were inoculated, one becoming infected. 
Parasites were intermittently present in scanty numbers in the one 
positive case. 

Case No. XI.—A goat aged one year, in poor condition and coming from a 
herd infected with scabies. July nth, inoculated subcutaneously with 5-0 c.c. 
blood from Case VIII. Organisms were not seen during the ensuing ten days. 
It arrived at the new camp on August 2nd, and trypanosomes were first seen on 
the 5th of that month. They continued intermittently present in scanty numbers 
for the six weeks of life remaining. 

The temperature, as in sheep, showed no tendency to reproduce the malarial 
type, but assumed rather that seen in the two infected cattle. 

This animal became badly infected with a form of dermatitis, which spread 
to the lips and around the eyes and ears, causing considerable irritation and 
probably hastened death, which occurred on September 13th, the sixty-fourth day 
after inoculation. 

Post-mortem .—Trypanosomes were not seen in blood or gland juice. Mucous 
membranes pale, a few petechiae on the conjunctivae. A few small circumscribed 
ulcers were present on the gums and hard-palate. No fluid in the thoracic and 
abdominal cavities. Heart pale and flabby, lungs normal. Liver pale but of firm 
consistence, capsule adherent to the diaphragm, gall bladder distended. In the 
large biliary canals several tape-worms were found. Spleen showed no gross 
alterations. Mesenteric glands pale and not greatly enlarged. Small concretions 
resembling those formed by (Esophagostomum columbianum occurred in the large 
intestine, (vide chart x.) 

Case No. XXXII. Healthy goat aged four months. August 7th, inoculated 
subcutaneously with i*o c.c. citrated blood of Case IX. The temperature was 
irregular, but trypanosomes were not seen, and it was alive and in good condition 
on November 20th. Sheep, Case XXXIII, inoculated at the same time became 
infected. 

Case No. XL. August 9th, inoculated with i*o c.c. blood direct from goat, 
Case II. The temperature showed a daily variation somewhat greater than usual, 
but trypanosomes were never seen. On September 17th it was inoculated with 
T. dimorphon and became infected on the twelfth day. Sheep, Case No. XXXIX, 
inoculated on August 9th from the same Case XI became infected. 

5. MONKEY. An adult female, probably Cercopithccus pygery- 
thrus , was inoculated subcutaneously on September 6th with roc.c. 
of blood from Case IX. It remained active and in perfect health 
until October 1 st, when it died suddenly through the maltreatment 
of the boy in charge. 

6. GuiNEA-PiGS. Five guinea-pigs were inoculated from, 
respectively, Cases IX, XI, XXXIII, XXXIX and X ( post-moriem ). 
They showed no disturbance in health, the temperature remained 
normal and organisms have never appeared. They are all still alive, 



ii 7 

February 18th, 1908, despite the rough usage to which they have been 
subjected on the march. 

7. DOGS. Three 1 kaffir 1 dogs were inoculated from Cases IX 
and XXXIII. The temperature remained normal, and organisms 
were never seen. One appeared to lose condition during the two 
months following inoculation, but later regained it, and with the 
others is now alive. 

8. Rabbit. A rabbit was inoculated on August 3rd with roc.c. 
of blood from Case IX, when parasites were present. Organisms 
were never seen, and it continued in perfect health until November 
14th, when it suddenly died. On post-mortem no signs of trypano¬ 
somiasis could be found. 

9. WHITE Rats. Five white rats were inoculated simultaneously 
with the five guinea-pigs. None have shown any disturbance in 
health, and four are still alive. The fifth was inoculated in November 
with a dog trypanosome, and died. 

INFECTION WITH T. THEIfJBRI 

During the routine blood examinations of cattle Nos. XVI, XXI 
and XXII, a large trypanosome was seen on one occasion in each. 
Further examinations in fresh and centrifuged specimens were 
negative. In Case XVI this trypanosome was seen three days after 
T . dimorphoti had been expelled by means of atoxyl, and four days 
before its reappearance. 

Case No. XXIII. A bull naturally infected with T. dimorfhon. August and. 
Inoculated subcutaneously with 90*0 c.c. citrated blood of Case XXII, which 
showed T. theileri that morning, T. dimorfhon also present. The animal died 
of T. dimorfhon infection three weeks later without ever showing T. theileri again. 

TRYPANOSOMIASIS IN SHEEP (T. DIMORPHON) 

A European who visited the camp informed us of mortality 
amongst his sheep, which were kept in a camp on the Lukanda river, 
45 miles North of Broken Hill. No specific details were obtainable, 
but it appears that between 30 and 40 had been sick and had died or 
been destroyed. Gl. morsitans exists all round, but infection by 
D. hepaticum was suspected. A visit was paid to this camp on 
November 23rd, during the owner’s absence, and three sheep found 
alive. One was in extremis , and blood examination revealed the 
presence of T. dimorphon. On post-mortem , there was an excess of 
fluid in all cavities, the lymphatic glands much swollen and oedematous 
but pale, the spleen was enlarged, soft and rounded. 

1 



n8 


NATURAL TRYPANOSOMIASIS IN DOGS 

We saw three dogs which were suspected by their owners of 
having naturally acquired trypanosomiasis. All had, at some recent 
date, been in the fly country around N’dola, 15 0 S., 24 0 40' E., and on 
examination showed trypanosomes and the clinical indications of the 
disease—a haggard appearance, anaemia, loss in condition, and in 
two, corneal opacity. 

Sub-inoculations were made from two of these dogs. 

From Dog ‘ A/ 

1. Guinea Pig. 

Case No. LXXII.—Nov. 8th, 1907, inoculated intraperitoneally with 1 c.cm. 
citrated blood. Parasites first seen on Nov. 20th and at each succeeding 
examination, about every fourth or fifth day, up till death on January 18th, 1908. 
Paralysis of the hind legs was noted on December 25th, but the animal recovered 
in three days. The same symptom reappeared on January 16th, and continued up 
till death two days later. Duration of disease 72 days. 

2. Rat. 

Case No. LXXIII.—Same date. Inoculated subcutaneously with i*o c.cm. 
citrated blood. Organisms appeared on November 18th (tenth day) and were 
present at each examination up to January 10th, when the animal died. Duration 
of disease 64 days. 

From Dog ' B.’ 

1. Guinea Pig. 

Case No. LXXVII.—November 15th, 1907, inoculated intraperitoneally with 
five drops of blood in citrate solution. Organisms appeared between November 
22nd and 26th, on which later date they were present in fair numbers. On one 
examination, made December 12th, they were not found but were present on all 
other occasions. Rat died January 12th. Duration 68 days. 

2. Rat. 

Case No. LXXVI.—Same date. Inoculated intraperitoneally with five drops 
of citrated blood. Parasites appeared between the 26th and 29th of November, and 
were constantly present on all succeeding examinations up to January 3rd, when 
it died—49 days. This rat had previously been inoculated with blood containing 
T. vivax y but had not shown infection. 


I. MORPHOLOGY OF CATTLE TRYPANOSOMES 
T. DIMORPHON 9 Dutton and Todd 

The parasite commonly encountered in the naturally infected and 
the experimentally inoculated cattle corresponds to the short forms of 
T. dimorphon described by Dutton and Todd as ‘ tadpole’ and 
f stumpy.’ The former predominated in all animals, except during 



the last few days of life in a few cases, when the stumpy type was 
more frequently seen. 

If the distinction between ' stumpy 1 and * long * is based upon the 
possession, by the latter, of a definite flagellum, this was not 
encountered in cattle. Its appearance, however, in sub-inoculations 
indicates that the parasite in question belongs to the dimorphon 
group. ' Tadpole ’ forms were found in every animal, with the 
exception of one moribund case mentioned later. They measured 
from 975 to 15*3 /4 in length, and up to 1*5 n in width. The body 
protoplasm stains a rather deep blue with the Giemsa stain, granules 
rarely being present; blepharoplast terminal or sub-terminal, small 
and rounded; nucleus a short oval, 17 to 2*5/4 in length, staining 
homogeneously a rather dark purple. Undulating membrane very 
rudimentary; in some, a small, fin-like, single fold could be seen. 
Flagellum absent, but an anterior prolongation of the protoplasm 
occasionally supported a minute extension of the rim of the 
undulating membrane. The posterior extremity is usually bluntly 
angular, but every gradation was met with. The greatest width lies 
posterior to the nucleus. Forms slightly larger than the above, which 
may be classed as * stumpy/ were occasionally seen in some animals, 
particularly in those in which the disease was running a more acute 
course, and have been constantly observed, but they become 
relatively more numerous towards death. In one animal, which had 
not shown organisms on two examinations previously, only this form 
was found at a third examination when the beast was moribund. It 
was not constantly to be observed at death; in five animals none but 
what are termed * tadpole * forms were seen. This form measured 
from 1775 to 21*25/4, and is of a stouter build than the 'tadpole/ 
The protoplasm assumes a pinker tint, the blepharoplast is better 
defined the nucleus almost invariably distinctly rounded and staining 
less deeply, and the widest part of the body is more usually on this 
level. An undulating membrane could be more easily distinguished, 
in some rare cases two or three folds being visible, and the bordering 
rim is commonly produced with the anterior extremity of the body to 
form an abruptly square-cut bristle-like beak, not exceeding 2/4 in 
length, but prominent in a well-stained preparation. Granules were 
seen in some, and divisional forms of this type were encountered. The 
distinction between this form and what is described as ' tadpole ’ is 



120 


more marked than between it and the ‘long’ form. These latter, 
that is to say, a trypanosome possessing a flagellum of more than a 
few fi in length, were not seen in cattle, and were only found in 
inoculated guinea-pigs and rats. In these animals they measured 
from 25 to 31 n in length, and could, on the whole, be clearly separated 
into the two arbitrary classes, ‘ male ’ and 1 female,’ the former being 
thinner, staining more homogeneously pink, the nucleus elongated, 
and the protoplasm free from granules; the latter stouter throughout 
the whole length, the nucleus oval, and the protoplasm taking a blue 
tint and often containing granules. The flagellum measured from 
5 to 11 /*, and continued from an undulating membrane which was not 
always so evident as in the larger of the ‘ stumpy ’ forms The 
blepharoplast is fairly prominent, rounded or oval, and commonly 
situated some little distance from the posterior extremity, which itself 
assumes most frequently a rather finely tapering point. The width 
of these forms varied between 15 and 2*5/1, dividing forms being as 
much as 3*5/1 across. While we refrain from the use of the expression 
‘ free flagellum,’ there appears little doubt that in the larger of the 
‘ long ’ forms, such a structure, free of cytoplasm, does exist. In the 
large 1 stumpy 1 and the short ‘ long,’ which cannot be clearly 
separated, the cytoplasm is unquestionably continued for a certain 
distance anterior to what we would regard as the normal extremity 
of the body, the place where body, undulating membrane and 
flagellum meet, and it is often impossible to determine the point at 
which this prolongation ceases. 

In fresh cover-glass preparations the corpuscular displacement is 
local and the organism does not readily pass out of the field of the 
microscope. The smaller forms cause, naturally, less commotion 
amongst the corpuscles. They are more readily retarded by these 
bodies, and the slow rolling displacement they produce is markedly 
different from the furious lashing of those with a long flagellum and 
the resulting scattering of the corpuscles. Neither form was ever 
noted to produce the peculiar ‘ catherine-wheel ’ effect seen in blood 
containing T. vivax. 

We were unable to make permanent slides from our donkey during 
the visit of November 20th, but Dr. Yale Massey kindly allowed us to 
examine the slides he had made on the day previous to the death of a 
naturally-infected donkey at Ruwe, in the Congo to the North of the 



I 2 I 


Rhodesian district we have under review. Short and long forms were 
present in this animal, the classification of 200 being * tadpole ’ 10 per 
cent., ‘ stumpy ’ 70 per cent., and 4 long ’ forms 20 per cent. The first- 
named measured between 10*5 and 1475/x, the ‘ stumpy * from 15 to 
18*25ft, and the ‘long’ forms between 18 and 27*25/1. In some of 
these latter, a flagellum measuring from 3 to 5 /i, which appeared quite 
free of cytoplasm, was seen. The structure of these several forms 
was essentially the same as that of those already given. 

In sheep and goats only the * tadpole ’ forms were seen. These 
measured from 9*75 to 14*5 in length and from i to 175/1 in width. 
In no particular could they be held to differ from those in bovine 
blood, and in the films examined only one form was seen which might 
possibly be regarded as ‘ stumpy.’ It has been noted that none of 
these animals died whilst under observation, and this may account 
for the non-detection of any but ‘ tadpole 1 forms. It is also to be 
remembered that in sheep, Case No. XXXIII, and goat, Case No. XL, 
the organism which appeared was of the ‘ tadpole ’ variety, whereas 
the guinea-pig from which they were inoculated had shown ‘ long ’ 
forms, and 1 stumpy ’ ones were present at the time of inoculation. In 
none of our inoculated dogs were forms longer than 16/1 seen. Most 
were of the ‘ tadpole ’ variety, but forms corresponding to ‘ stumpy ’ 
were encountered towards the end of the disease. We reserve the 
description of the organisms encountered in the naturally infected 
animals till later. 

In rabbits, in the films made from our single case, only * tadpole * 
forms were seen. 

Guinea-pigs are the most satisfactory animals for revealing the 
dimorphic variations of this trypanosome. It has been noted that the 
average period of duration of organisms was ten days. During the 
first four days, ' tadpole ’ forms are almost exclusively present, giving 
place between the fifth and eighth days to f stumpy ’ and ‘ long *; 
whilst at death, and on the two or three days previous, ‘ stumpy * forms 
predominate. 

The 1 tadpole ’ and ‘ stumpy * forms correspond to the descriptions 
given, while that of the 4 long ’ form is based on its appearance in these 
animals. We do not hesitate to say that in many of these, 
particularly the so-called male forms, a flagellum whose length 
sometimes exceeded 10/* was seen free from cytoplasm. These forms 



122 


were present, though less numerous than the ‘ stumpy/ in blood 
inoculated into the goat, Case No. XXXIII, sheep XL, and rat 
LXXI, in which the ‘ tadpole ’ forms were reproduced. 

Rats were infected by blood containing all forms. 1 Tadpole ’ 
forms predominated throughout the first few days of the disease, 
while the ‘ stumpy ’ became more numerous towards death. * Long ’ 
forms were very rare, but were seen in those inoculated from an ox, 
a goat, and a guinea-pig; but they were not seen after inoculation 
from a dog or a second goat infection. It is to be remarked, 
however, that a thorough examination of all slides has not been 
possible. 

We have here, then, a trypanosome whose prevailing type in 
naturally and experimentally infected animals is short, measuring 
only 975 to 15’3>“, and from which there is a relative absence of a 
flagellum. This type under the influence of a different host, or under 
natural conditions in the same animal, assumes a distinct form which 
measures from 25 to 31 n in length, and possesses a flagellum which 
may be upwards of 10 jjl long. With the knowledge at present 
available there can be no • hesitation in ascribing the name 
Trypanosoma dimorphon , Dutton and Todd, 1904, to such a 
dimorphic organism. 


II. T. ViVAX, Ziemann 

We have already remarked that the movement of this organism in 
fresh cover-glass preparation is of extraordinary rapidity, and the 
effect produced by the passage across any one field closely resembles 
that of a spirochaete, the corpuscular displacement being transitory 
and of no greater magnitude than that produced by drawing a floating 
hair across the surface of still water. It is impossible to retain any 
one organism in the field by movement of the slide, even when the 
mechanical stage is not employed, the parasite crossing and altering 
its direction with bewildering swiftness. In a preparation which has 
been kept for an hour or two the movements of some become more 
sluggish, and in fields where corpuscles are scanty, progression is seen 
to be due to a rotatory motion of the whole body, to the exclusion of 
a wavy undulating membrane and the vibrations or lashings of a free 
flagellum. 



When one of these parasites is obstructed by corpuscles or fibrin 
threads, the action again differs from that of other trypanosomes. 
Sometimes the ordinary lashings are produced, but more frequently 
the posterior end becoming a fixed point, the anterior circles round 
with regular sweeps, forcing the corpuscles away as the arc-like 
radiations pass from a burning catherine-wheel. The trypanosomes 
measure from 20 to 26/1 in length, and up to 3*4// at the widest part, 
posterior to the nucleus. The body tapers anteriorly from this in a 
rather regular fashion, while the posterior end is distinctly rounded. 
The blepharoplast is large, round or oval, measuring up to I'/i in 
diameter, and is usually terminal. The nucleus is commonly an 
elongate oval, up to 375/1 in length and 2*5/x in breadth (average 
3 4 by 2), and occupies nearly the whole transverse diameter at this 
level. The undulating membrane is very narrow, about 1 /1, the 
bordering rim arises from the neighbourhood of the blepharoplast and 
is continued as the flagellum after running parallel to the body, so 
giving an aspect of stiffness to the whole structure. For the greater 
part of its length this 4 free ’ flagellum is accompanied by a 
continuation of the periplast, which may or may not contain cytoplasm. 
The actual free extremity does not appear more than 3*5 /1 in length, 
commonly less; whilst the total length of the tapering end anterior 
to the cessation of the membrane is upwards of 8*5/1. The length of 
this whip was somewhat greater in the later stages of the disease, thus 
our figures in the inoculated calf, case IX, on August 4th (twenty- 
fourth day after inoculation) give an average of 3*4/1 ; those on 
September 8th (fifty-eighth day) vary between 3*4 and 8*5, with an 
average of 5*9/1. 

The cytoplasm stains homogeneously; vacuoles are seldom seen 
and granules are not very common. Dividing forms are rare ; in the 
peripheral blood we have only seen those with a double blepharoplast 
We have never noted the alveolar arrangement of the cytoplasm 
described and figured by Liihe. 6 This trypanosome has retained its 
quite characteristic movement in fresh preparations and the same 
appearance in stained films, excepting only the slight differences in 
the length of the anterior extremity, in all animals which took the 
experimental infection. 



124 


III. THE TRYPANOSOMES OF NATURALLY-INFECTED DOGS 

The forms seen in all three animals are the same. The trypano¬ 
somes measure from 20*25 to 28*9/* in length, and from 1*5 to 
2*5 fi in width. The protoplasm stains pink with Giemsa, vacuoles 
and granules were inconstant; the posterior extremity is most 
commonly pointed. The nucleus is situated towards the anterior 
part of the body, and varies in shape from a round to an elongate oval, 
the former taking a deeper stain than the latter. The blepharoplast 
is rounded, usually terminal, but may be removed from the posterior 
extremity by as much as 2*5 /*. A well-developed undulating 
membrane is present, and the rim is continued as a flagellum, which 
varies from 3 to 10// in length, and is accompanied, in the short forms 
at least, by a prolongation of the cytoplasm. Those carrying a short 
flagellum correspond in type to the so-called ‘ female ’ forms, and the 
long flagellar forms to the * male' of other trypanosomes. It would 
be impossible to assert that these ‘ females ’ are not the ‘ stumpy ’ 
forms of T. dimorpkon , which are somewhat larger than normal. 
Trypanosomes which could in any way be considered as having any 
relation to 4 tadpoles ’ were not encountered; the smallest measure¬ 
ment made was 20*25/1. In the two guinea-pigs, the trypanosomes 
which reappeared measured from 21 to 30/1 ; in the smaller forms 
the projecting flagellum is short. The variations in length noted 
depend mainly upon the size of this flagellum; the body itself being 
fairly constant at 18 to 20/1. Tadpole-like forms were not seen at 
any stage of the infection. 

In rats no forms except 1 long ’ ones were met with. These 
measured from 21 to 30/1, and from 1*5 to 2/1 in width. There is a 
distinct flagellar appendage in all, which differ from those seen in 
guinea-pigs only in being more granular and staining less deeply. 

The natural dog trypanosomes are monomorphic in so far as no 
variations comparable to those existing between ‘ tadpole ’ and 4 long ’ 
forms were seen. Being monomorphic and of large size, a parasite 
of a domestic animal and occurring in Africa, this trypanosome must 
be placed in that heterogeneous collection whose type is T. brucei. 



125 


ARGUMENT 

It must be admitted that our present knowledge of trypanosomes 
does not permit of any satisfactory classification. In Africa the 
confusion in nomenclature is appalling, and the number of specific 
or suggested names, based largely upon the country of origin or the 
first found host rather than upon morphological or biological 
characteristics, renders absolute diagnosis of an individual form 
almost impossible without a typical living strain for comparison. 

There is perhaps no great difficulty in asserting that the body 
structure of certain trypanosomes shows dimorphic variations under 
certain conditions. In all trypanosomes morphological differences 
between individuals occur to a greater or lesser extent, but only in the 
one species, T . dimorphon , Dutton and* Todd, are they considerable. 
In this, unless the ‘ tadpole ’ and the ‘ long 1 be seen, they are not 
striking; and between the larger of the ‘ stumpy ’ and the shorter of 
the * long ’ we do not consider the variations greater than between 
some of the so-cailed ‘ male ’ and ‘ female,’ or the smaller or the larger 
forms of J. evansi. An additional difficulty is that there is no one 
structural point that can be seized upon as a basis for classification; 
most gradations can be met with in the shape of the posterior 
extremity and the length of the anterior and the amount of the 
flagellum that is free. As we have found forms recognisable as 
‘ tadpoles ’ and ‘ long ’ both under natural conditions and those of 
ordinary animal experimentation, we have no hesitation in asserting 
that the cattle trypanosome first described is dimorphic according to 
the interpretation placed upon that word by workers on trypano¬ 
somiasis. 

The first dimorphic trypanosome was that described in the Gambia 
by Dutton and Todd, 1 who at the same time described the patho¬ 
logical reactions produced by it there, which reactions were later 
confirmed in Europe by Thomas and Breinl, 2 and by Laveran and 
Mesnil. 3 They showed that rats, guinea-pigs and, with one 
exception, dogs are susceptible, and die within one or two months. 
The trypanosome encountered in the cattle of the Congo Free State 
was also dimorphic in type, but the animal reactions in that country 
differ from those obtained in the Gambia. At one post, Romee, 
laboratory animals were shown to be highly susceptible ; at the other 



126 


posts twenty-nine inoculations were made from cattle whose blood 
showed trypanosomes of this type. One rat showed organisms twice, 
after the original and after a re-inoculation, but for one day only in 
each case ; a second rat only became infected after several months’ 
incubation. Dutton, Todd and Kinghom 4 consider this Congo 
trypanosome as T. dimorphon on the grounds that the morphological 
characteristics of this species are peculiar in the genus Trypanosoma , 
and identical with those of the form they describe; and that the 
variation in virulence is not sufficient proof that more than one species 
of trypanosomes was present. The trypanosome which we describe 
shows animal reactions approaching those of the Gambian strain ; all 
our dogs, rats and guinea-pigs have succumbed to an acute infection 
within two months, and, as in Gambia, the rabbit has a more chronic 
disease and the cattle an acute form. With similar morphological 
appearances and similar animal reactions, we consider the Rhodesian 
form to be T. dimorphon , Dutton and Todd, 1904. 

We have much more hesitation in ascribing the second of the 
Rhodesian cattle trypanosomes to a specific class. The morphological 
characteristics seen in animals which were susceptible are not 
sufficient to consider it dimorphic; ‘ tadpole ’ forms, or any approach 
to these, were not encountered; the variations seen were not greater 
than have been found in monomorphic trypanosomes, and to this 
group it is assigned. We distinguish it from T. dimorphon which 
occurred in cattle of the same herds and also in some of the same 
animals by: - 

1. The morphological appearances; the fairly constant size 

and the extreme rapidity. 

2. The clinical type of disease induced; the great daily varia¬ 

tion of temperature as opposed to the ‘ curve ’ in cattle 
suffering from T. dimorphon y and the absence of the 
striking febrile reactions seen in sheep and goats. 

3. The animal reactions: Five rats, five guinea-pigs and three 

dogs were inoculated without effect One goat and three 
sheep did not become infected, and in those which did 
there were indications of recovery. In one goat and one 
sheep which had been inoculated with this trypanosome, 
and were not showing parasites, a re-inoculation with 



127 


T. dtmorphon brought about a febrile reaction and the 
appearance of this organism in the blood. 

4. The post-mortem appearances: Splenic enlargement was 
not noted in the three cases upon which autopsies were 
possible, whereas in T . dimorphon infection it was a 
common feature. 

Sander and Hennig 6 state that according to Ziemann T. vivax 
occurs spontaneously in cattle, sheep and goats ; the incubation period 
is 5-8 days; and on post-mortem , enlargement of the liver or spleen 
is seen. Experimentally, grey rats died in 8-11 days; in donkeys 
the disease is chronic, and in a German dog the reaction was 
apparently doubtful. A negative result was obtained in a white rat. 
In Rhodesia white rats and native dogs were negative; the one 
donkey inoculated did not show organisms, but it has not been 
examined or reported on for three months and we are ignorant of its 
present state. A point of difference we note is the enlargement of 
the spleen, but our observations are based on three post-mortems 
only. 

The morphology, as given by Liihe, differs from that of the form 
under discussion in possessing an alveolar protoplasm and a somewhat 
pointed end. These are minor points, negligible so far as the 
morphology of other trypanosomes is concerned, in which similar 
variations are commonly seen. The experimental work on both sides 
is limited, but the animals used are similar, and if Ziemann’s dog was 
negative, and such is a possible interpretation, the results coincide, 
though we were unable to use grey rats and pigs in which Ziemann 
obtained positive reactions. 

We know of no other trypanosome whose activity approaches 
that of this form. On morphological grounds, then, and animal 
reactions this trypanosome coincides more closely to that of Ziemann 
than to any other, and with all reserve, we feel justified in considering 
the second parasite of Rhodesian cattle a$ sufficiently closely allied to 
T. vivax , Ziemann, to bear that name until the classification of the 
genus be put on a more satisfactory basis. 



128 


TRANSMISSION OF CATTLE TRYPANOSOMES 

European and native unite in incriminating the tsetse-fly common 
throughout the Northern part of the area of North-Western Rhodesia. 
The examinations of our specimens of these has so far only shown 
Glossina morsitans , the approximate distribution of which is marked 
on the attached map ; but it must be understood that the lack of signs 
in certain areas does not imply freedom from the fly, but only that we 
have no positive knowledge of its occurrence. 

Tabanidae have only been incriminated in the one instance 
already recorded. During June, July and August, 1907, we did not 
see any, but during the latter end of September they were 
occasionally seen at our camp, and they were very numerous in 
Broken Hill on the first few days of October. On our line of march 
they were constantly encountered, and may be held as having an 
almost universal distribution. The larger members of this family are 
locally known as ‘ hippo flies ’; Haematapota , also common in 
November and December, are usually spoken of as ‘ blind flies.’ 

Stomoxys were taken in the cattle kraal of the farm where our 
camp was established in July and August, and they were caught on 
the River Kafue in November and on the River Luapula in December. 
They were most frequently met with in villages, but on two occasions 
were taken from recently shot game. 

Lyperosia were caught in the same cattle kraal in July and the 
first week in August, and again during the latter part of September. 
They were not seen in the interval, nor have they been taken any¬ 
where on the route followed. 

Hippoboscidae are, in comparison with India, rare. One specimen 
of H. rufipes (?) was shown us as coming from near the Kafue. We 
have thrice taken Lipoptena on dead buck. 

Owing to the relative frequency of Glossina and their association 
with game and domestic animals, there can be no question that, 
assuming game to be the pre-existing reservoir, they are the most 
capable flies for transmitting the trypanosome to cattle or other 
domestic animals taken into their haunts. We may mention that we 
have seen trypanosomes in fresh preparations of blood made from a 
recently shot Hartebeest (Bubalis lichsiensteini ) and a bush buck 
(Tragelaphus scriptus ), but our examination of the slides and 
inoculated animals is incomplete. 



129 


Nearly all the cases of trypanosomiasis in cattle examined could 
be given a history, often very imperfect, of having at some recent date 
been exposed to the tsetse; but in one herd, where the history is 
reliable, the evidence is suggestive that Stomoxys and Lyperosia had 
acted as transmitting agents. 

Forty-four animals from this herd had lived at Kapopo, which 
itself is free from fly, for upwards of three years, during which time 
they had always been in good health and deaths were rare. In 
August, 1906, they were brought to Broken Hill, a distance of about 
97 miles, following as far as possible a route where tsetse were scanty 
or not known to exist, and adopting the usual precautions, such as 
marching at night. They remained in perfect health until June, 1907, 
when they came under observation. During these eleven months two 
animals had died, apparently of some acute inflammatory disease, and 
one had been destroyed as the result of an accident, and six others 
which had not been recently in tsetse areas were added. The farm 
where these cattle were kept is at least two and a half miles from the 
nearest known fly area, and they were all employed on this farm in 
agricultural work ; the three cows and three bulls grazing close to the 
buildings. On April 18th, six bullocks were sent on a journey of 
thirty miles Southwards to Mwomboshi, and returned four days later. 
G. morsitans occurs on the road travelled about eight miles from the 
farm, but it is limited to a narrow patch. On June 25th, three of 
these animals and one which had not been away showed trypano¬ 
somes, and they were all dead within a month; one died within ten 
days, and this at the height of the dry season, when deaths are said 
not to take place. Four other animals were suspected, but did not 
then show organisms. The rest of the herd were all in good condition, 
looking bright and doing the hard agricultural work well. On July 
18th, all remaining animals were examined, and eleven, some of them 
straight from the plough, showed trypanosomes. The owner, who 
possessed a small microscope, picked out another on the 22nd, one on 
the 27th, and we found two more on the 29th. Three of these were 
cows which had not been exposed to Glossina for twelve months at 
least. 

All these animals were segregated, and those not showing 
organisms were placed by themselves in a kraal and grazing area 
which appeared free of all biting flies, such as Stomoxys. and 



130 


Lyperosia. Further examination of these apparently healthy 
animals weeded out five more cases in August. The balance, fourteen, 
continued healthy, and of these thirteen were taken for transport 
work in September. They did the work well, and showed no greater 
death-rate on the 400-mile march than did the other ninety cattle 
taken. 

After the first examination, at which only suspected cattle were 
presented, all animals inspected were, without exception, free of any 
clinical signs of the disease. These became manifest in about two 
weeks time, and death took place within an average of thirty days of 
diagnosis. This is the average between these dates in fourteen 
apparently healthy cattle, and excludes all those upon which experi¬ 
mental work on treatment could be held as influencing the course of 
the disease in either direction. 

With a disease of such rapidity and virulence, we consider it 
highly improbable for the infection to have lain dormant in the 
animals since August, 1906. From an examination of all conditions, 
we think it probable that one or more of the six cattle which went to 
Mwomboshi in April contracted the disease on the road and brought 
it to the farm, where, in the presence of Stomoxys and Lyperosia in 
the kraals, these animals, including cows and bulls, which did not 
leave the place, became infected, and that the segregation from these 
flies checked its spread to the fourteen cattle which remained healthy. 
The one animal which did not go on trek with the thirteen cited 
above was brought to live with the sick on September 10th. It was 
then in excellent condition, but on a visit to the farm on November 
20th it showed T. dimorphon , and would in all probability succumb 
within two weeks. Owing to the lack of facilities, and the pressure of 
other work, we were unable to conduct any transmission experiments 
with these two flies, but as affording corroborative evidence we 
examined the road on which the outbreak was suspected of having 
originated. We were informed by the owners concerned that they 
had lost animals during the present year after travelling through this 
area with their spans, and this despite the usual precautions. We, 
therefore, exposed two healthy bullocks to Glossina morsitans there. 

Cases No. XLIV, XLV.—Two animals from small herd at Mwomboshi, where 
they had lived for between two and three years, and where stock apparently does 
well. August 19th, 1907. These two were driven between 10 a.m. and 1 p.m. 
over the road, through the area inhabited by Gl. morsitans. Only three flies were 



seen to feed, two on No. XLV, and one on No. XLIV. On arrival at our camp 
they were segregated and carefully kept from all association with other cattle. 
The temperature remained normal until September 3rd, the fifteenth day after 
being bitten. On September 6th, the eighteenth day, trypanosomes were seen in 
the blood of both. This trypanosome was morphologically identical with that we 
have called T. vivax. On September 13th (XLV] and September 16th (XLIV) 
T. dimorfhon appeared, and both organisms w£re present until September 30th, 
when No. XLV died. Both had been submitted to treatment by Atoxyl and 
Mercury, and No. XLIV has been detailed elsewhere, (vide chart ii.) 

This double infection by such a small number of flies as were seen, 
and on a road where game is scarce, caused further enquiries to be 
made, and we found that a span of oxen had travelled that road on 
August 14th, five days prior to the date our animals passed. On 
September 21st we examined this span, and found two animals 
infected with T. dimorphon , and a third showing both T . dimorphon 
and T. vivax. 

If the Glossina mors it an s which bit Cases Nos. XLIV and XLV 
had not derived these organisms from game, it would appear that 
they have the power of transmitting five days after the infecting feed, 
which would have been taken from this span, whose owner reported 
that he had lost several animals during the months of June to 
September from what he regarded as ‘ fly ’ infection. 


CONCLUSIONS 

1. That trypanosomiasis of domestic stock is very prevalent in 
the Northern area of North-Western Rhodesia, and that it is due to 
T . dimorphon (Dutton and Todd), T. vivax (Ziemann), and one 
morphologically allied to T. brucei (Plimmer and Bradford). 
T. theileri also occurs, but does not appear to cause serious damage. 

2. That these trypanosomes may be transmitted by Glossina 
morsitans , Stomoxys calcitrans , and a species of Lyperosia . In 
nature it will depend upon the conditions under which cattle are 
maintained, to which of these genera special attention must be paid 
in prophylaxis. 

3. That treatment, as detailed in a previous report, shows certain 
indications of success, and we urge facilities for a continuation of the 
work on this disease, which tends to stagnate the proper development 
of a wealthy mineral and agricultural country. 



IJ 2 


REFERENCES 

1. Dutton and Todd. The Liverpool School of Tropical Medicine, Memoir XI. 

2. Thomas H. Wolferstan and Breinl, A. The Liverpool School of Tropical 

Medicine, Memoir XVI. 

3. Lave ran and Mesnil. Trypanosomes et Trypanosomiases, Paris, 1904. 

4. Dutton, Todd, and Kinghorn. The Annals of Tropical Medicine aud 

Parasitology. Vol. 1, No 2, pp. 233-271. 

5. Luhe. Mense, Handbuch der Tropenkrankheiten, Leipzig 1906, Band III, I, 

P 125. 

6. Sander and Hennig, idem, Band III, II, p. 721. 








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133 


REPORT ON THE WORK OF THE GREEK 
ANTIMALARIA LEAGUE DURING THE 

YEAR 1907 

BY 

M. HADJIMICHALIS, President 

AND 

JEAN P. CARDAMATIS, General Secretary 
(Received for publication 13 April , 1908) 

I 

Greece takes rank among those countries which are the most 
infested by malaria. 

This plague existed here even in the remotest periods of antiquity, 
and Hippocrates, the father of medicine, not only mentions in his 
works all the various forms of the disease, but he also was aware 
of its connection with marshes, and of the influence of certain 
meteorological conditions, especially the frequency of rain, upon its 
development. 

We know that malaria has always existed in Greece since that 
time, and Professor Ross is certainly correct in attributing to a great 
extent the cause of her misfortunes, to this dreadful scourge. 

Official statistical information, concerning this disease, exists only 
for the last nine years, and is limited to the 12 largest towns of 
Greece, with a population of over 10,000 inhabitants each, or a total 
of 446,743 souls. 

According to these statistics (Table A) it is shewn that the 
average annual number of deaths from malaria, in the 12 towns 
referred to, is 287 or 9-8 per 10,000 inhabitants. 

The first place, as regards the number of deaths, is held by Volo, 
in Thessaly (21*89 P er 10,000); next in order follow Pyrgos, in the 
Peloponnesus (19*48), Larissa (17) and Triccala, in Thessaly, (1472) 
and Calamata, in the Peloponnesus (13*38). 

It must, however, be observed, that these figures do not give an 
exact idea of the prevalence of malaria throughout the country, as 
K 



134 

this disease is much less frequent in the towns than in the rural 
districts. 

That this is the case is clearly shewn by the information which 
has been collected by the League, and in the publications of many 
physicians. In fact malaria is so widely spread in our country that 
scarcely any communities are free from the disease. 

The plains of Thessaly, Phthiotis, Acamania, Boeotia, Elis, 
Messenia, Argos and Laconia are all severely scourged by this 
plague, and, at certain periods, hardly a single inhabitant of those 
districts escapes the disease. 

For instance, an examination of the school children, held at 
Marathon in October, 1906, shewed that enlargement of the spleen 
was to be found in every pupil (100 per cent.). 

The disease usually begins in the month of May, reaches its 
height in July and August, and commences disappearing in November, 
but in the more elevated districts it appears later and disappears 
earlier. However, even in winter relapses are very frequent. 

Further statistical information may be found in a small work, 
published by Professor C. Savas (Le paludisme en Grece et l’oeuvre 
de la Ligue Antimalarienne. Atti della Societa per gli Studi della 
malaria, tome VIII, p. 139, 1907). 


II 

The year 1907 was signalised by a severe epidemic of malaria in 
Greece, as was shewn by the private information which reached the 
League from all parts of the country, as well as by official statistics 
(Table A). 

As regards, however, the number of cases of the disease, the year 
1907 fell short of 1905, whilst the intervening year, 1906, was 
characterized by a considerable decline in this respect. 

In 1907 the greatest number of deaths from malaria took place in 
July (Table B), August shewing the second highest results, but whilst 
in the previous years June and September shewed the highest figures, 
on the contrary, in the year under review a slight decrease in the 
mortality from the disease was observable in September, followed 
immediately by a fresh increase in October, which was maintained 
throughout November. This curious fact, which was observed for 



135 


the first time during this year, is to be attributed to the prevailing 
atmospheric conditions, as, after a warm summer, lasting until the 
middle of August, the temperature fell in September, after which it 
again commenced to rise and remained relatively high during October 
and November. 

Consequently, the disease, which had commenced to abate in 
September, began to re-assert itself in October, and continued in 
this way throughout November, and even, to a certain extent, in 
December. 

The records, as regards fatal cases of malaria in the year 1907, 
were held by the Thessalian towns, especially Volo (with 45 deaths 
per 10,000 inhabitants) followed by Triccala (24) and Larissa (23). 
The second rank was filled by two Peloponnesian towns; Pyrgos (15 
deaths per 10,000), and Calamata (13). 

The capital, Athens, itself was severely afflicted by this plague 
during the year under examination, for, whilst the average annual 
number of deaths from malaria in that city (as shewn in Table A) is 
56, during 1907 no fewer than 71 persons died of the disease, or, in 
other words, that year was second only to the year 1901 in respect of 
the number of fatal cases of malaria. 

The cause of this prevalence was found to be in the stagnant pools 
remaining in the river-bed of the Ilissus. The draining of the Ilissus 
was commenced some three years ago, and those quarters of the city, 
bordering on the drained portion of the river-bed, were found to have 
been but slightly affected by the disease this year, whilst the portion 
of the city adjoining the section of the river where the draining 
operations bad not yet been completed, were severely attacked. 

Ill 

One of the chief cares of the Antimalarian League, after its 
establishment three years ago, was the collection of information 
concerning the propagation of malaria. 

In view of the fact that the official health statistics are confined 
entirely to the publication of the number of deaths from various 
diseases in the 12 largest towns of the kingdom, the League 
endeavoured to supply this deficiency as far as possible, and with this 
object a printed circular was addressed to all the physicians with the 



136 


request that they would fill in their answers to the questions therein 
asked. This was also done in 1907. At the same time a bulky 
volume was issued, containing all the material accumulated by the 
League during the years 1905 and 1906 (together with the information 
gleaned by the Panhellenic Congress of 1901) consisting of the 
reports of 450 physicians on malaria in three hundred demes of the 
country. 

It is hoped that the Statistics of the remaining 120 demes will 
soon reach us (as we are quite without information as to the progress 
of the disease in these demes), and the malaria chart of the Kingdom 
will thus be completed. 

This volume in addition contains the transactions of the League 
during the two years 1905, 1906, together with a number of scientific 
articles, and was issued in an edition of 3,000 copies and distributed 
gratis to all the physicians of the Kingdom, so as to serve as an 
incentive to their aid in the common struggle against malaria. 

We also sfcnt 200 copies to the medical men of Crete, accom¬ 
panied by a circular requesting information respecting the disease in 
their island during the year 1907. 

The High Commissioner of Crete, Mr. A. Zaimis, was pleased to 
evince the greatest interest in the suppression of malaria, which 
infests that island, and he gave the necessary instructions for the 
commencement of a methodical campaign against the disease. 

The Councillor of the Interior also promised to do all in his power 
to assist in the campaign in view of the progress which has already 
been made in Greece. 

Before proceeding to more important work, the League consi¬ 
dered it better to obtain the advice of some of the more eminent 
provincial medical men, who are actually engaged in the field of 
battle, and who would be able, by their experience and valuable 
advice, to afford us considerable assistance. 

The summoning of these physicians to Athens would be another 
considerable advantage to the League in its work, as they would be 
thus enabled to come into closer contact with us, even on a short visit 
only, and would be able to discuss the newest ideas, in connection 
with the propagation and suppression of malaria, and, on their return 
to their own districts, they would be able to act as apostles of the 
League and to endeavour to instil the principles of the Leagu^ by 



137 


means of lectures, &c., and in other ways to further the aims of the 
League. 

Inspired by these ideas, the Committee decided to invite to 
Athens, at the expense of the League, a number of these medical 
men, and, on the 3rd and 23rd of May O.S. two medical meetings 
were held and were attended by about 50 physicians from the 
different provinces of the Kingdom. 

Each meeting lasted only two days, and the best means of com¬ 
bating the malarial disease was discussed. With the aid of the 
lantern and the microscope, in the Laboratories of the Hygienic and 
Pathologic Anatomical Institute, the malaria parasites and the 
various kinds of mosquitoes were shewn upon the screen, whilst the 
causes of malaria and the means of protection against the disease 
were explained at length in accordance with the most up-to-date 
theories. Visits were further paid to the bed of the Ilissus river and 
the breeding places of the Anophelines in the stagnant water. 

We consider that the assembly of these doctors at Athens met 
with complete success, and that it will conduce to the dissemination of 
the work of the League, as they have already formed, in their own 
districts, centres for the inculcation of the ideas of the League among 
the other local medical men. 

It gives us great pleasure to mention that only three of the doctors 
attending the Congress asked for, and received, their travelling and 
hotel expenses. 

The President has received letters from some of these gentlemen, 
shewing that they are indeed making great efforts to realise the aims 
of the League, as they have both delivered lectures and have 
approached the local municipal Councils with proposals for the voting 
of funds, besides collecting money themselves for the purpose of 
draining the pools in the neighbourhood of the various communities. 

The action of the Board was not limited to the convocation of the 
physicians, but, actuated by the desire to place the information con¬ 
cerning the means of combating the disease before as wide a circle 
as possible, so that the people might be furnished with the most 
up-to-date ideas, the League printed and distributed gratis 30,000 
copies of a pamphlet containing information regarding the propa¬ 
gation of malaria and instruction as to the means of prevention. 

In addition, the General Secretary of the League, Dr. J. 



138 


Cardamatis, was dispatched to 30 towns of the Kingdom in order to 
deliver lectures on the same subjects, which were attended by a large 
number of people, including the medical men and local authorities. 

By means of these lectures, together with the distribution of 
printed instructions, and the indication of the anopheline mosquitoes, 
the interest of all was aroused with regard to this vital question. 

The expenditure incurred in connection with the meetings and the 
lecturing tour of the General Secretary, amounted to Drs. 1.917.55 
(about £70). The most important work undertaken by the League 
in the year 1907, was to combat malaria at Marathon. 

The League considered that lectures and publications, and 
similar measures were not sufficient to achieve its eminently practical 
aims, but that practical application of the measures recommended 
by the League was also necessary for the persuasion of the public 
as to the efficacy of the remedies recommended, whilst, on the other 
hand, a precedent should be established for the execution of similar 
works in the future. 

This advice had also been given by the Liverpool §chool of 
Tropical Medicine in its most valuable report on the suggested anti- 
malarial measures in Greece, dated the 25th of January, 1907. 

The League, however, had other reasons for considering this work 
to be necessary, as we ourselves should study, in practice, the con¬ 
ditions under which the combating of malaria in Greece was 
possible, taking into consideration the social and local peculiarities of 
the country. With this object the Committee selected Marathon as 
the field of action of the League. 

In this respect Marathon presents many advantages, as it is, in 
the first place, one of the districts which suffers most from malaria; 
secondly, the plain of Marathon is separated from the surrounding 
districts by a range of hills, so that it forms a completely independent 
region; and finally, it lies at a convenient distance from Athens, so 
that the work could be carried on under the perpetual surveyance of 
the League. The sole disadvantage connected with the choice of 
that district lay in the fact that the number of inhabitants is rather 
large for the purpose (1,680 souls) and is scattered throughout the 
plain, thus rendering the expenses rather heavier than would other¬ 
wise be the case. 

The work was commenced on April the 27th, by the establishment 



*30 


in the village of a permanent Ambulance consisting of one doctor 
with a medical student as assistant, whilst the General Secretary of 
the League remained there alternately with the assistant of the 
Bacteriological Laboratory at the University, so that the Staff always 
consisted of three members, under the general direction of Professor 
Savas and Dr. Cardamatis. 

The plain of Marathon lies at a distance of 3O kilometres from 
Athens, and contains three villages, Marathon, the capital of the 
deme, with 1,200 inhabitants, Bey, with 150, and Souli, with 160 
inhabitants. There are, in addition to these villages, several small 
hamlets scattered here and there in the plain, inhabited by 173 souls 
in all, giving a total population of the plain of 1,680. 

According to the statements of the local doctors, the number of 
cases of malaria averages between 80 per cent, and go per cent, of 
the population. Dr. Papasotirios examined, during the month of 
October, 1906, the spleen of all the pupils of both the schools, and 
found them enlarged in the case of 100 per cent. 

Besides this, of 1,216 individuals, whom we examined in the 
month of May O.S., 1,031 or 85 per cent, admitted that they had 
suffered from marsh fever during the previous summer. This large 
number of cases of malaria is chiefly due to the neighbouring river¬ 
bed, the waters of which decrease in volume in the summer months, 
and leave pools full of larvae of Anopheles superpictns. 

The peasants are also inoculated with the disease during the 
night, when sleeping in their vineyards, which are situated in the 
plain, close to several pools and two large marshes, in which breeding- 
places of Anophelines were always discovered, and especially 
Anopheles claviger , superpictiis and less often bifurcatus . 

The work was commenced in two ways, first by ridding the waters 
of the river-bed of the Anophelines, and, secondly, by the regular 
distribution of quinine to all the inhabitants as a curative and 
preventive measure. 

The waters of the river-bed were concentrated in a narrow 
channel, in order to assure a rapid flow, and the pools were covered 
with petroleum once a week. 

This work was partly carried out by workmen, but chiefly by the 
pupils of the schools, who gladly assisted under the leadership of their 
teacher, and they were thus afforded an opportunity of a practical 



140 


lesson as to the manner of communication of the malaria, and the 
means of safeguarding themselves against the disease. 

The second method of prevention consisted in the distribution of 
quinine. Owing to the difficulty of supervising the inhabitants as 
regards the use of quinine, consequent on the manner in which they 
are scattered throughout the plain, and to the insufficiency of the 
medical staff, we preferred giving out the quinine according to the 
Koch’s system, which in practice was slightly modified, e.g., the drug 
was distributed on both Saturdays and Sundays, in doses of 1 gramme 
each day. We must, however, confess that we often met with 
considerable difficulty in the application of this mode of distribution, 
as the giving of a comparatively large quantity at once caused much 
inconvenience to the people, hindering them in their work, or 
rendering their Sunday’s rest burdensome. 

During the whole of the six months of our work, we distributed 
23 kilogrammes of quinine and 1 kilogramme of euchinine. The 
euchinine and 5 kilogrammes of sulphate of quinine were presented 
to us by the Jobst-Zimmer firm, whilst the Italian Government was 
kind enough to send us 3.100 kilogrammes (3 1/10) of the State 
quinine, prepared in the manner in which it is sold by that Govern¬ 
ment in Italy. 

The latter method of putting up quinine was very much appre¬ 
ciated by the peasants, and the children, especially, readily took the 
tannate of quinine with chocolate. 

Our experience with the euchinine was not so favourable, as it 
proved to be much less readily taken than the tannate with chocolate, 
and less efficacious. 

The quinine provided by ourselves was the sulphate, and was 
distributed in wafer covers. 

Of the 1,680 inhabitants of Marathon 1,544 underwent the treat¬ 
ment, but we are without information as to the result of the cure as 
regards many of these people. Of 1,252 persons, however, we possess 
the necessary information, and of these only 597, or 47*6 per cent., 
were attacked by the disease. 

A more detailed examination of these figures shews that of 67 
persons whp took quinine for 21 to 24 weeks, none were attacked by 
malaria. Of 145 who took the drug for 16 to 20 weeks, 36 suffered, 
or 20*6 per cent. Of 220 who took quinine during 11 to 16 weeks, 



103 were attacked, or 48*6 per cent.; whilst of 820, who took quinine 
irregularly and for periods from 1 to 10 weeks, 464 were attacked by 
malaria, or 56*5 per cent. 

It should be here noted that in the surrounding villages, as well 
as in the whole of Attica, the malaria was very severe during the 
period under review. 

The average amount of quinine consumed by each inhabitant, 
undergoing the preventive and curative treatment, was 15*6 grammes. 

With regard to fatal cases of malaria, in the village of Marathon 
there died during the summer of 1907 one child of.one year old, of 
pernicious spasmodic fever, and one girl of seventeen years of age, of 
blackwater fever. In the village of Bey, a child, aged 4 years, died 
of the latter disease. None of these children had undergone our 
preventive treatment. We have no information as to the number of 
deaths from malaria in the summer of the year 1906, but in 1905, 
seven died of that disease in the village of Marathon. 

We hope that we shall be able to continue our work at Marathon 
under better auspices in the coming summer, when we shall have the 
advantage of the experience hitherto acquired. 

The whole expenditure incurred in the six months* work at 
Marathon, amounted to Drs. 5.715.65 (£210), of which salary and 
travelling expense of the doctors, rent and sundry expenses connected 
with the staff, accounted for Drs. 3.548.70. The purchase of quinine, 
drugs, &c., Drs. 1.897.25, Petroleum and sundries Drs. 239.20. 

The amount expended averages Drs. 3.70 per head for the six 
months, or less than that expended by the Italian Red Cross Society 
(which in 1901 amounted to fr. 11.25 per head) and the Austrian 
Government (Kr. 9.50 in 1903 and 1904). 

Besides the sum expended at Marathon and the expenses on the 
two medical Congresses and the lecturing tour, a further amount of 
Drs. 6.820 (£250) was accounted for by the printing of the above- 
mentioned Statistics of the League, the detailed instructions 
regarding the prevention of malaria, and of the under-mentioned 
appeal on the part of the League. 

The funds, which rendered possible the work of the League, were 
supplied by philanthropy. 

The Committee formed in England, on the initiative of Professor 
Dr. Ross, under the presidency of Sir Alfred Jones, and which H.R.H. 



I 4 2 


Princess Christian graciously condescended to favour with her 
patronage, contributed a sum of £740, collected in Egypt and 
England. 

Notwithstanding the fact that the thanks of the League were 
conveyed at the time by the President to the English Committee, we 
take this opportunity of again expressing our deep gratitude to all 
those friends, who, through their subscriptions so largely contributed 
to the success of the campaign, the chief object of which is to free 
our country from this age-long scourge, which has during so many 
centuries been the source of incalculable harm to the Greek nation. 

Before concluding this Report, we must add a few words in con¬ 
nection with the Bill concerning quinine, which was drawn up and 
submitted to the Government two years ago through the instru¬ 
mentality of the League. 

In view of the fact that the quinine offered for sale, in the 
remoter parts of the Kingdom, is> not only often of bad quality, but 
is also sold at a high price, and is further taken without any method 
by the peasants (who, owing to the frequency of the attacks of the 
disease, do not always consult a doctor, but treat themselves), the 
League drew up and submitted to the Chamber a bill providing for 
the undertaking of the sale of quinine by the Government. 

In order that the Government, the Press, and Public opinion in 
general should be fully informed on the subject, the League issued 
and distributed 6,000 copies of an appeal (in March, 1907) in which 
it exposed the harm done by malaria, and, at the same time pointed 
out the remedies against the evil, chief amongst which were the 
draining of the pools and small marshes in the neighbourhood of 
dwellings and the introduction of a monopoly of quinine. 

In this appeal there were published at the same time : (1) a report 
on the combating of malaria in Greece, sent us at our request by the 
Liverpool School of Tropical Medecine; (2) the Bill concerning 
quinine, with the report relating thereto, and, (3) a translation of the 
Italian laws concerning the combating of malaria. 

The Committee of the Chamber, to which the Bill was forwarded, 
effected certain alterations therein, in conformity with the Italian 
system of selling quinine, and the amended Bill was passed in its last 
reading on December 15th of last year, becoming a law of the 
Kingdom. 



x 43 


According to this law, the Government has the right to procure 
and to sell any salt of quinine, which may be designated by the Board 
of Health. The quinine will be ordered through the Ministry of 
Finance on the basis of tenders (except if it is purchased from a 
foreign Government) on terms to be arranged on each occasion by 
the Board of Health. The limit for each order is fixed at two years. 

The sale of quinine will be effected through the Chemical 
Laboratory of the Ministry of Finance, the Public Treasury, the Post 
and Telegraph Office, the Public School Teachers and by other public 
Offices, in virtue of a Royal decree. 

The State will sell quinine at cost price, and a reasonable profit 
will be allowed to retailers. In the case of the sulphate and 
bisulphate of quinine, the law fixes the price per gramme for retail 
sale at a maximum of io lepta. The prices of other salts will be 
fixed by Royal decree as occasion arises. 

This law does not prohibit the free import and sale of quinine, 
but quinine so imported will be chemically analysed before its entry 
into the country. 

Penalties are appointed for the sale of State or other quinine at 
a higher price than that fixed, for adulteration of or the sale of 
adulterated quinine, or for smuggling quinine into the country, as well 
as for selling the article under weight. 

The law further obliges those demes which suffer severely from 
malaria to enter in their budget an amount sufficient for the purchase 
of State quinine for the free supply to the indigent. 

The above is a succinct account of the action of the League during 
the past year. As regards the future, the first item in the programme 
is the resumption of the campaign at Marathon for a series of years, 
and, subsequently, by the convocation of more doctors at the capital, 
and by means of lectures in other towns of the Kingdom, to con¬ 
tribute to the dissemination of the latest ideas regarding the pre¬ 
vention of malaria, and, to sum up, by means of suitable action with 
the Government and the municipalities together with the large landed 
proprietors and the public in general, to aim at the application of 
measures for the drainage of the large number of large and small 
marshes which cover our country. 

This difficult work, however, imperatively calls for the co-operation 
of philanthropy. 



T 44 

























TABLE B. Number of Deaths in each Month during the Year 1907 


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147 


A PECULIAR INTRALOBULAR CIR¬ 
RHOSIS OF THE LIVER PRODUCED 
BY THE PROTOZOAL PARASITE OF 
KALA-AZAR 


RY 

LEONARD ROGERS, M.D., F.R.C.P., B.S., F.R.C.S., I.M.S. 

PROFESSOR OF PATHOLOGY, MF.DICAI. COLLEGE, CALCUTTA 


(Received for publication 16 March , 1908) 


In my report on Kala-azar, published in 1897, I described a 
dropsical form of the disease, which appeared, in part at least, to be 
produced by cirrhosis of the liver, which was found in one case post 
mortem. More recent experience has shown this complication to be 
commoner in the more chronic sporadic form of kala-azar, which is 
so frequent in Calcutta, than it was in the Assam epidemic disease. 
These cases, however, were very difficult to verify by finding the 
parasite by spleen puncture on account of the organ receding into the 
abdominal fluid before the point of the needle, although the character¬ 
istic history, greatly enlarged spleen and leucopenia, left no doubt 
in the mind of an experienced physician regarding the true nature 
of the cases. Some degree of fibrosis of the liver was described in 
my first 1897 report, while about a year ago a post mortem on a 
chronic case of kala-azar, which died with all the classical symptoms 
of cirrhosis of the liver, has enabled me to study the advanced stages 
of the affection. I, therefore, propose in the present paper to give a 
somewhat fuller account of the affection than the note in my work on 
Fevers in the Tropics, in which the photograph of a kala-azar patient 
suffering from this type of cirrhosis of the liver is given (opposite 
page 67). The following are the notes of the case referred to above. 

L 



i 4 8 


CHRONIC SPORADIC KALA-AZAR TERMINATING WITH 
CIRRHOSIS OF THE LIVER 

The patient was a Hindu male, aged about 30. He had suffered 
from enlarged spleen with frequent attacks of fever on and off for 
five or six years. During the last six months he had taken country 
liquor, but not in excess. He was in hospital for 35 days before his 
death, during which time he suffered from ascites with enlargement 
of the liver and spleen, and persistent diarrhoea, and was greatly 
emaciated and anaemic, and had a troublesome cough. No malarial 
parasites could be found in his blood, and he only occasionally had 
a slight rise of temperature. The disease was diagnosed as 
kala-azar. 

A post mortem was performed and the following conditions noted. 
The body was extremely emaciated, and the subcutaneous, fat was 
very scanty. The peritoneal cavity contained 30 ozs. of clear fluid, 
but there was no oedema of the legs, the dropsy having decreased 
with the diarrhoea while in hospital. The pleural cavities showed 
some fibrous adhesions, but the lungs were crepitant throughout, 
although slightly congested at the bases. Both the parietal and 
visceral pericardium showed extensive haemorrhages (these being 
common in kala-azar in various positions). The heart muscle was 
somewhat pale, but firm. The stomach showed petechial haemor¬ 
rhages in the mucous membrane. The small intestines showed a few 
points of haemorrhage due to the bites of a small number of anchylos- 
tomata (such as are found in 75 per cent, of post mortems in natives 
in Calcutta), but were quite free from ulceration. The large intestine 
showed some pigmented scars, while the mucous membrane of the 
lower part was congested and oedematous. The liver weighed 
29 ozs. (the body weight was only 60 lbs.). The surface was perfectly 
smooth, and of a greenish brown colour. It cut very firmly, and its 
substance could not be broken down by very firm digital pressure. 
The gall bladder was healthy. The spleen weighed 12^ ozs., 
having become much diminished in size during the continuance of 
the diarrhoea. The capsule was wrinkled, and on section it was a 
brownish-red colour and very firm, owing to excess of fibrous tissue. 
The kidneys were healthy except that the capsule stripped with some 
difficulty, but left a smooth surface. The brain was healthy. 



149 


MICROSCOPICAL EXAMINATION 

The protozoal parasites of kala-azar were found in large numbers 
in the bone marrow, spleen and liver, being mostly of the small size 
seen in chronic cases of the disease. In the liver they were found 
with an oil immersion lens in the endothelial cells of the capillaries 
between the columns of hepatic cells in specially prepared specimens. 
With haematoxylin and eosin only very fine dots could be detected 
in this position, being the nuclei of the parasites, closely resembling 
pigmentation, for which I think I must have sometimes mistaken 
them in my original Assam investigation in 1896. The persistence 
of the parasites in the advanced cirrhotic stage of the organ is 
remarkable, and leaves little doubt that they are the cause of this 
peculiar intralobular cirrhosis. 

The general appearance of the liver under a low magnification 
(Zeiss A. Oc. 2) is shown in fig. 1 of Plate I. To the left is seen 
a portion of the capsule at the site of an extensive fibrous band, but 
it will be observed that there is no marked depression of the surface 
such as produces the hobnail appearance of the common atrophic 
cirrhosis of the liver. The capsule itself also shows but very slight 
thickening. The most striking feature is the universal distribution 
of the cirrhotic process throughout the liver lobules, so that the hyper¬ 
plastic connective tissue widely separates each column of epithelial 
liver cells, and, indeed, makes up the greater bulk of the lobules from 
the portal to the hepatic venules. A careful study of a number of 
sections showed that there is extremely little alteration in the general 
arrangement of the liver lobules, which retain to a great extent their 
shape and size, although the intralobular veins are somewhat less 
prominent than usual. There is distinct cellular and fibrous increase 
around the portal interlobular veins, but not extending far round the 
circumference of the liver lobules as a rule, so that the organ is not 
cut up into small areas of hepatic substance by complete circles of 
fibrous tissue, as in atrophic cirrhosis. This explains the absence 
of the typical yellow lobulated appearance to the naked eye on 
section of the organ, which is so characteristic of hobnail liver, and 
has probably led to the new form having been frequently over¬ 
looked, especially in its less marked degrees. The perilobular portal 
tissue shows a few well-marked bile-duct-like double columns of some- 



150 


what cubical epithelial cells, but these are nothing like so numerous 
as in hypertrophic cirrhosis of the liver, being, indeed, scarcely more 
evident than in the atrophic form. 

Fig. 2 of Plate I shows a portion of a liver lobule under a higher 
magnification (Zeiss A, Oc. 2), and includes some of a periportal 
fibrous band. The very great increase of the intercellular connective 
tissue of the lobule will be at once apparent, the epithelial cells 
forming barely half of the area of the lobule, each column of liver 
cells being separated from the next by an extensive layer of fibro- 
cellular connective tissue. In places the liver cells contained much 
yellow pigment derived from broken down red cells anaemia being 
a marked symptom of the later stages of kala-azar, although fre¬ 
quently only slight in degree in the first few months of the disease, 
much less so than in true malarial fever of any duration. With this 
exception the surviving liver cells have a fairly healthy appearance, 
and stain well. The connective tissue between the liver cells is 
partly fibrous, but chiefly consists of small round cells together with 
a considerable number of larger epitheloid-like ones, some of consi¬ 
derable size. It is the latter which were found to still contain the 
parasites of kala-azar in specially stained sections, and they are, 
doubtless the enlarged endothelial cells of the capillary vessels, 
which S. R. Christophers first described as containing the human 
stage of the kala-azar parasite. 

This distribution of the organism at once furnishes the key to the 
peculiar position of the connective tissue proliferation. These 
minute protozoa multiply in the endothelial cells, and on reaching 
their full size some of the cells rupture, scattering the parasites into 
the blood stream, where they are found in comparatively small 
numbers, in the polynuclear leucocytes more particularly. These 
bring them back again to the liver, spleen and bone marrow, where 
the cycle is repeated. When this process continues for a number of 
years, as in the chronic form in which alone I have seen cirrhosis of 
the liver supervene, it is not surprising that eventually the constant 
irritation of the parasite causes proliferation of the connective tissue 
around the capillary vessels throughout the liver lobules, and to a 
less extent in Glisson s capsule, around the portal radicles, and so 
produces the condition above described. This universal thickening 
round the capillary vessels of the liver causes both an extensive 



atrophy of the hepatic cells and also must considerably retard the 
circulation of the portal blood. It thus produces the marked ascites, 
which, although the fluid accumulates much less rapidly than in 
atrophic cirrhosis, yet is a formidable and not infrequently fatal 
complication in these unfortunate people, worn out as they are by 
years of fever, and reduced to an extremely debilitated and emaciated 
state. 


FREQUENCY OF INTRALOBULAR CIRRHOSIS OF THE 
LIVER IN KALA-AZAR 

Among 48 post mortems I have performed on sporadic kala-azar 
in Calcutta in the last few years, marked cirrhotic changes were 
present in the liver in four; while in seven more, slighter degrees of 
fibrosis were met with. The latter number is certainly too low, for 
in half the cases the exact consistence of the organ was not recorded 
in the post mortem notes, and this degree is very easily overlooked. 
On the other hand, the liver was noted to be softer than normal in 
nine cases, so even slight fibrosis is very far from being constantly 
met with. This is due to the extreme variation in the duration of 
this fever, namely, from a few months to five to ten years. In my 
work on Fevers in the Tropics a table of the degree of enlargement 
of the liver in different stages of kala-azar is given, from which it 
appears that marked enlargement of the liver is rarely seen before 
the end of six months fever, while the cirrhotic condition usually only 
appears after several years illness. 

MALARIAL CIRRHOSIS OF THE LIVER 

In view of the fact that until the last few years kala-azar has 
always been classed as ‘ malarial cachexia,’ the discovery of the 
above-described form of cirrhosis due to kala-azar raises the question 
as to how far descriptions of malarial cirrhosis of the liver may have 
been based on cases of kala-azar erroneously diagnosed as malarial. 
In this connection it is worth recording that in five years pathological 
experience at the Medical College, Calcutta, I have only once met 
with a case of undoubted malarial cirrhosis of the liver, in which the 
microscopical picture of uniform extensive thickening of the peri¬ 
lobular connective tissue with much black pigment in its lymph 
spaces, and to a less extent in the intracellular tissue throughout the 



15*2 

liver lobules, was so characteristic that it could not possibly be over¬ 
looked My impression, therefore, is that a true malarial cirrhosis 
of the liver does occur, but that it is certainly decidedly rare even 
in highly malarious Lower Bengal, for a large proportion of our cases 
come from the unhealthy districts surrounding Calcutta. 

I may also mention that typical atrophic cirrhosis is extremely 
common in Bengal, more so even than in Europe, although it is 
certainly not as a rule due to alcohol. Major O. W. Sutherland has 
also recorded a similar experience in the Punjab. Thus in five per 
cent, of over 4,000 post mortems in Calcutta cirrhosis of the liver was 
found, although 40 per cent, of the deaths were from typically tropical 
diseases, such as cholera, &c. If these are excluded the percentage 
rises to between 8 and 9 per cent, of the deaths. On the other hand, 
in Berlin Forster found cirrhosis of the liver in but I per cent, of 
3,200 post mortems. This very important subject, however, is 
beyond the scope of the present paper. 


SUMMARY 

1. The most chronic cases of kala-azar not infrequently terminate 
their course with ascites due to cirrhosis of the liver. 

2. The cirrhosis is of a peculiar intralobular type of uniform 
distribution and with a smooth surface to the organ. 

3. It is due to the protozoal parasite of kala-azar, which may be 
found in the liver and other organs after death. 

4. This form of cirrhosis of the liver is much commoner in Lower 
Bengal than a true malarial cirrhosis, with which it has probably 
hitherto been confused. It is, however, much less common than 
atrophic cirrhosis due to unknown causes. 



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153 


WHAT IS ‘SCHISTOSOMUM MAN SONF 

Sambon 1907? 

BY 

Dr. A. LOOSS, 

PROKK.SSOR UK PARASITOI.CH'.Y, SCHOOL OK MEDICINE, CAIRO 

{Received for publication 21 March , 1908) 

About a year ago Dr. SAMBON, of the London School of Tropical 
Medicine, startled the scientific world interested in human para¬ 
sitology by the creation of a new species of blood fluke, Schistosomum 
mansoniy which he stated had hitherto been confounded with Sch. 
luzmatobium (1907a, p. 117). A suggestion to this effect had been 
made as far back as 1903 by Sir PATRICK M ANSON. SAMBON’s 
new species was thus readily adopted by MANSON in the new edition 
of his * Tropical Diseases* (1907, p. 660). From the West Indies 
there had come information which seemed to corroborate Dr. 
SAMBON’s views (HOLCOMB, 1907). Later, the author gave a some¬ 
what fuller account of the new species (1907b, p. 303), and quite 
recently, he mentioned its existence as a fact in a paper ‘ On the 
Part played by Metazoan Parasites in Tropical Pathology/ read 
before the Society of Tropical Medicine and Hygiene, London (1908, 
p. 29Q. In this paper and in the ensuing discussion repeated allusion 
was made to the views hitherto held with regard to Bilharzia and 
Bilharziosis by the workers in Egypt in general, and particularly by 
myself. It was hinted that we had not recognised the differences 
between two easily distinguishable species. If Dr. SAMBON’s view 
were correct, all of us who have devoted attention to the subject, 
would have indeed been wandering in the dark since the time of 
BlLHARZ himself, fifty-seven years ago. Since such a charge has 
practically been made I feel it necessary' to take up the defence. 
I may as well at once say that when in London two years ago I 
dropped some well-meant hints of warning to be cautious, whether 
to Sir Patrick Manson or to Dr. Sambon I do not remember. I 
am sorry that these hints have not been heeded, for if so the present 
disagreeable discussion might not have become necessary. 



154 


Being at present fully occupied with some other work I must 
limit myself to the discussion of some points of primary importance; 
but it is possible that I may take an early opportunity of returning 
to the subject in more detail. Speaking quite generally, I may say 
even now that those of us who have seen anything of Bilharziosis in 
Egypt are convinced that the scientific problem it offers is more 
complicated than Dr. SAMBON seems to imagine; it is a problem 
which will require a long and close collaboration of the Anatomist, 
the Pathologist, and the Helminthologist before it may be considered 
as solved in every detail. There is, further, one thing which can not 
be too strongly emphasised at the very outset, and this is that any 
theory, be it ever so cleverly based on the biology of the parasite, 
must be wrong if it contradicts the facts supplied by the Anatomist 
and the Pathologist; and also that any other theory, however 
plausible its explanation of the anatomo-pathological observations, 
can not represent the truth so long as it is irreconcilable in any detail 
with the biology of the parasite. I may mention in passing that 
several theories recently brought forward as explanations of the 
aetiology of certain human diseases caused by worms are open to the 
latter objection; the theory of SAMBON is the latest of these, and a 
very interesting one it is, not perhaps so much for the arguments by 
which, but on account of the manner in which it is supported. 

In order to make the purport of what I have to say hereafter 
quite clear, I will state that I do not consider it my task to prove 
whether or no there exists a Schistosomum mansoni. Among 
scientific workers it is a good custom that anyone who believes he 
has made a new discovery also takes the trouble to prove it; it is 
not customary among scientists to assert something and call for the 
help of others to establish it. In the case which I am about to discuss, 
it is Dr. SAMBON who, acting upon a suggestion of Sir PATRICK 
MANSON, formally published Schistosomum mansoni as a new species. 
After the usage generally adopted in science, the merit of the 
discovery is his when the discovery is right. But with him also must 
rest the responsibility of bringing forward all the evidence which may 
be reasonably demanded in support of it. Dr. SAMBON, indeed, 
supports his action by a certain amount of evidence; but he 
is obviously aware himself that, especially, its zoological part 
(i.e. the possibility of distinguishing the adult forms) is 



*55 


practically nil. He therefore concludes his answer to the various 
objections made in the discussion by expressing the hope (in a form, 
by the way, which I fail to appreciate) that I would soon be able to 
provide that description of the adult forms which he himself was 
unable to produce (p. 46). I am sorry that I cannot accept the part 
thus assigned to me. If Dr. SAMBON had not sufficient material to 
demonstrate beyond doubt the specific independence of Sch. mansoni , 
he might with advantage have postponed the publication of the name 
until such necessary material was available. Since he has gone so 
far as to publish the name, and thereby implicitly claims to have 
made an important discovery, I think that it is incumbent upon him, 
and not upon me (or any other), to do the work of supplying such 
proof as the rest of the scientific world may ask. As I have already 
s^id, I cannot consider it as my legitimate task to prove or disprove 
the existence of a ‘ Sch. mansoni .’ What I propose to do is to point 
out the inadequacies of Dr. SAMBON’s theory. Doing this, i.e., 
giving the reasons against SAMBON’s theory, amounts to 
practically the same thing as giving the reasons for the views held 
by me with regard to some fundamental items in the biology of 
Sch. hcematobium. I am not displeased to have this opportunity: for 
the rest, every reader is free to form his own judgment. 

The reasons which lead Dr. SAMBON to assert the existence of a 
separate species, ‘ Sch. mansonil are three: a zoological, a patho¬ 
logical, and a geographical one. The first is afforded ‘ chiefly ’ by the 
ova. ‘ In Sch. hcematobium the eggs are more or less lanceolate, and 
provided with a short, straight, terminal spine; in Sch. bovis they are 
spindle-shaped, and provided with a short, terminal, heart-shaped 
spine; in Sch. japonicum they are ovoid, and have no spine; and in 
Sch. mansoni they are oval and provided with a stout, lateral spine * 
(1908, p. 31). The adults producing the two varieties of eggs are as 
yet indistinguishable. Dr. SAMBON ‘ had the opportunity of 
examining several specimens collected at post mortems in Egypt and 
Uganda.’ He ‘ noticed that whilst the majority of female worms 
contained within their uterine tubes the characteristic ova of 
Sch. hcematobium , with a short terminal spine at the posterior 
extremity, two presented lateral-spined ova. These had been 
removed from the gynaecophoric canal of males differing in no 
appreciable way from those clasping the more common kind. 



Unfortunately, the material at hand was so badly preserved that it 
precluded any study of comparative anatomy ’ (1907b, p. 303). 

The second specific character of Sch. mansoni is, according to 
Dr. SAMBON, given in its ‘ different anatomical habitat * and its 
‘specific pathogenic action* (1908, p. 32). ‘ Sch. mansoni does not 

affect the genito urinary organs, its ova are eliminated solely by way 
of the intestine ; they are never found in the urine. The patients 
harbouring this parasite suffer from a haemorrhagic enteritis, but 
they never present haematuria ’ (1907a, p. 117). The third reason for 
assuming the existence of a Schistosomum different from Sch. 
hcematobium , is found by Dr. SAMBON in the peculiar geographical 
distribution of Sch. mansoni. According to the data published in 
literature, the new species is ‘ probably ’ alone present in the West 
Indies, for endemic haematuria is unknown there. The same may be 
said with regard to the Congo Free State, where careful recent 
investigations have shown the absence of haematuric bilharziosis and 
the frequency of a rectal infection, in which the ova of the parasite 
bear invariably a lateral spine. In the Cape Colony, op the other 
hand, haematuria is very common; HARLEY, Brock, and others 
working in those districts state in their articles on the subject, that 
they never encountered the egg with the lateral spine. In Egypt, 
both Sch. hcematobium and Sch. mansoni are found side by side, but 
the former appears to be far more prevalent, and is certainly more 
in evidence, owing to the haematuria to which it gives rise. That is 
probably the reason why the two forms have been confounded, and 
the scarce, laterally-spined ova looked upon as abnormal and 
distorted (p. 32). SIR PATRICK MANSON, on page 660 of his text¬ 
book, shortly states that BlLHARZ in 1851 noted the presence of a 
Schistosomum producing lateral-spined eggs, but confounded it with 
Sch. haematobium. 

We will now analyse these reasons given by Dr. SAMBON some¬ 
what in detail. I begin with the zoological. It is the most 
important; for the foundation of a new species is, to put it briefly, in 
the first place a ‘ zoological act.’ In order to establish a new species 
safely it is necessary to point out constantly present 
and, if possible, easily recognisable zoological 
characters by which it may be distinguished from related 
forms. The less constant and the less definite the characters of a 



presumed new species are, the more it is contestable from the 
zoological standpoint. The characters themselves must, first and 
foremost, be derived from the adult stages. With regard to 
this point, Dr. Sambon’s evidence is nil. He has examined some 
badly preserved specimens, but the males showed no difference at all 
from those of Sch. heematobium , and the females differed in the shape 
of the egg only. To this latter we shall return later; speaking of 
the adults, I may state, from a general point of view, that I would 
not, a priori , consider it as a serious objection to Dr. SAMBON’s views 
if there really were no marked anatomical differences between the 
adults of the supposed two species. There are some few cases known in 
which certain forms resemble each other to such an extent that they 
might well be representatives of the same species, did not other 
factors—such as they are known to us at present seemingly exclude 
the possibility of the forms being the same thing. 
Dr. SAMBON, in order to make the absence of all distinctive 
characters of his new species appear less weighty, dwells at some 
length on two cases in which, after a long and tedious comparison of 
many adult specimens, I have myself come to similar conclusions. It 
would lead me too far to discuss these cases in detail here. I will only 
remark that one of them has no bearing on the case at present under 
discussion, inasmuch as the forms in question show differences 
which, though slight, are yet sufficiently pronounced to enable any 
expert to distinguish the respective forms as easily as he may 
distinguish Sch. heematobium and Sch. bovis. In the second case, a 
parasite was found to inhabit several mammals, but to be entirely 
absent from birds, in Europe; whereas a similar form, in North 
Africa, could never be found in the same mammals, but was present 
in birds which never visit Europe. In this case, I of course depend 
upon the facts available at present, and it is very probable that a 
comparison of a larger supply of new material (my own investigations 
were made 15 years ago) will reveal structural differences here also. 
But unless the difference be cleared up by new observations I feel 
compelled to consider the respective forms as different, in spite of 
their apparent structural identity. However, Dr. SAMBON, or any¬ 
body else, is fully at liberty to show that the premises on which my 
opinion is based are erroneous. If he succeeds in showing this by 
irrefutable facts I shall certainly be the first to change my opinion. 



158 

But, before doing so, I want to hear facts, just as in the present 
article I am about to point out a number of facts which are 
irreconcilable with Dr. Sambon’s theory. He assures his audience 
that * certainly there were more and better reasons to separate Sch. 
wansoni from Sch. hcematobium ’ (1908b, p. 46) than there were in my 
two cases referred to by him. We shall see how far this is true. 

Dr. SAMBON first pretends that the other known Schistosoma 
species do not show any marked differences in their adult stages. 

‘ The Sch. bovis, for instance, resembles the Sch. hcematobium so 
closely that, indeed, it would be very difficult for anyone to point out 
any marked difference between the adult forms of the cattle parasite 
and those of Sch. hcematobium ’ (1908b, p. 46). To this I have to 
reply that Dr. SAMBON is mistaken. To ‘ anyone * who actually has 
some helminthological knowledge a single glance with the naked eye 
will suffice to tell Sch. hcematobium from Sch. bovis , and a good 
pocket lens will suffice to differentiate Sch. japonicum from Sch. 
hcematobium. There are, in addition, quite well-marked internal 
differences which Dr. SAMBON might have known had he consulted 
the latest description given of Sch. bovis by Leuckart (1894, p. 470!), 
or the short review 1 gave of KATSURADA’s paper on Sch. japonicum 
(1905a). The fact that Dr. SAMBON is not apparently aware of the 
existence of these differences is in itself a poor reason for the state¬ 
ment to his audience that they are really absent. As a matter of 
fact, in no case does the differentiation of any of the species of 
Schistosommn hitherto described, whether affecting man or animals, 
depend on the form of the egg alone. Since the various Schistosomes 
affecting animals are not mentioned by Dr. SAMBON, I will not refer 
to them any further in this discussion. 

Thus, Dr. SAMBON is not able to produce any distinctive anatomi¬ 
cal character of the adult Sch. mansoni. There remains only the 
egg. It is a well-known fact that man y, but by no means a 11, 
species of parasites may be recognised from their eggs. If this is 
the case, the aspect of the egg is one of the distinctive characters 
of certain species. I do not, however, at present remember one 
single case in which two species of parasitic worms acknow¬ 
ledged as independent differed solely by their eggs. 
The fact is easily comprehensible. If I cannot tell whether two 
specimens I have before me are individuals of one species or 



159 


individuals of two species, I cannot tell either whether slight 
differences I observe in their eggs are specific characters or not. If 
I so desire, I may assert that there are two species; but, in that case, 
others will certainly demand proofs of such a statement. Dr. Sam BON 
pretends that the two shapes of the egg found in association with 
Sch. hamatobium belong to two different species, but I cannot see 
that he can possibly prove this zoologically without finding distinc¬ 
tive differences between the adults. For the proof must consist in 
showing that one form of egg is constantly connected with a certain 
anatomical structure, and the other form as constantly connected with 
another anatomical structure of the adults. Until this is done I am 
afraid that Sch. mansoni will find little approval with zoologists, 
in spite of Dr. SAMBON’s contention that ‘ to zoologists the characters 
of the ovum should suffice for the determination of a new species ’ 
(1908a, p. 31). 

The remarkable difference in the position of the spine of the egg 
of Sch. hczmatobium has long attracted the attention of observers, the 
majority of whom considered the egg with the end spine as the 
normal, and that with the side spine as abnormal. Various attempts 
have been made to explain the formation of the latter. Dr. SAMBON 
refers to these theories, but in a rather peculiar manner. He parti¬ 
cularly mentions FRITSCH, ‘who had described certain differences in 
the genital tract of the female, but was under the impression that the 
females containing the lateral-spined ova belonged to the same 
species as those containing terminal-spined ova. He therefore 
explained the difference by abnormality. FRiTSCH’s explanation was 
obviously wrong, but his description was perfectly correct 1 (1908b, p. 
46). I should like to know in what way Dr. SAMBON has obtained 
the evidence for the concluding part of this statement. He has said 
that the two females he had an opportunity of examining were so 
badly preserved that any study of their anatomy was precluded. 
How, then, does Dr. SAMBON know that Fritsch’s description was 
1 perfectly correct ’ ? I doubt whether he has at all read that author’s 
original article (it is, unfortunately, not accessible to me at present); 
he has certainly not read the later descriptions of LORTET and 
VlALLETON, LEUCKART and myself, in the latter two of which 
FRITSCH’s statements with regard to the point under discussion are 
refuted as incorrect. 



i6o 


I will not tire the reader by a long anatomical description of the 
structures at issue; suffice it to state that up to this day I have 
personally found only one type in the structure of the internal 
genital organs of the female, although the uterus may contain in o n e 
specimen ova with a terminal, and in another specimen ova 
with a lateral spine. The position of the spine does not depend upon 
a preformed difference in the internal structure (which, of course, 
changes its shape somewhat with the contractions of the body), but 
on the relative position of the egg during the process of its formation 
in the ootype. I have tried to show this in a diagrammatic drawing 
which has recently been copied in various books on Bilharziosis; I 
may mention in passing that in this figure the lateral-spined egg is 
placed unusually steep; I have in the meantime come across worms 
in which the axis of the egg lay almost at right angles to the axis of 
the ootype. Dr. SAMBON ignores the existence of this drawing as 
well as the descriptions of LEUCKART and myself; I should like to 
submit that he will have to account for them if he wants to maintain 
Sch. mansoni as an independent species. 

On the whole, the zoological characters of the new species are as 
vague as they can possibly be. Dr. SAMBON is himself aware of that 
and refers to a case where, in one instance, ornithologists have 
based a new species solely upon the character of the egg. I am not 
in a position to criticise the actions of ornithologists; but the 
fact that they find something justified is for me not in itself a reason 
to consider the same thing as justifiable also in helminthology. I 
would mention, by the way, that the new species of bird will 
certainly not be generally accepted unless it can be shown that the 
aberrant shape of the egg is reasonably constant. 

The details thus far mentioned are in the main of a technical 
zoological nature. I should not have been compelled to enter upon 
them had not Dr. SAMBON tried to show that the foundation of Sch. 
mansoni was justifiable from the zoological standpoint. That it 
cannot be, will become obvious even to the non-specialist by another 
fact not mentioned by Dr. SAMBON. The fact is that in Egypt, 
the eggs of Sch. hcematobium and 'Sch. mansoni ’ 
may occur in one and the same individual 
female. This observation is now 57 years old and might have 
been known to Dr. SAMBON, had he studied the papers of those 



i6i 

authors whom he accuses of having failed to recognise an obvious 
fact The observation is due to BlLHARZ. It is true that BlLHARZ 
did not yet know how to interpret those bodies which we now describe 
as lateral-spined eggs; but this is of no importance as compared with 
the fact, that once he found one of these enigmatical bodies in the 
anterior part of the uterus of a female, the posterior part of which 
was filled with the ordinary ova. That there was no mistake 
possible may be gathered from the circumstance that BlLHARZ, on 
a later occasion, and after having discovered the same bodies in the 
tissues of the liver and the rectum, emphatically repeats that 'such 
a body was, though once only, but quite un¬ 
doubtedly, found in the uterus of a female 
worm, the posterior part of which contained 
the normal ova’ (BlLHARZ, 1852, pp. 74 and 75). Besides, 
BlLHARZ has proved too careful an observer to admit of any mistake 
on his part; as a matter of fact, many a recent ‘ discovery ’ with 
regard to Bilharzia and Bilharziosis may be found described in his 
paper when one takes the trouble to read it. 

If my memory does not quite fail me, I have in the course of 
years, myself seen several similar females; but considering the 
occurrence of both shapes of eggs in the same individual as anything 
but new, and not foreseeing either the importance the specimens 
would one day gain, I have not separated them from the rest, and it 
is quite possible that one or the other may be found in the material 
which I have sent away from here to various places. I very much 
regret that at the present moment I cannot produce a specimen. It is 
a curious fact, of which we shall have to speak again later, that the 
portal veins very often contain only males; the worms within recent 
years found at the post mortems in the Kasr el Aini Hospital, and kindly 
left to me by Dr. FERGUSON, were almost exclusively males; in one 
of the last cases, e.g., there were 64 males but not a single female. 
I have, however, no doubt that sooner or later I shall be able to 
establish the accuracy of BlLHARZ’s observation by the production 
of an actual specimen. 

The occurrence of terminal-spined and latejal-spined eggs in one 
and the same individual worm is one of the fundamental facts on 
which my views rest; I wonder how Dr. SAMBON will explain it by 
his theory. 



162 


I have said above that, a priori, a great structural similarity of the 
adult stages would not necessarily be a proof of there being only one 
species. If, on the other hand, I am asked to acknowledge a specific 
difference between Sch. hamatobium and Sck. mansoni , in spite 
of their great internal resemblance, I certainly expect that the 
other proofs in favour of the existence of a separate species will 
be absolutely clear and stringent. We will now see how these parts 
of Dr. Sambon’s evidence stand an earnest scientific test. 

There are numerous cases where closely allied parasites (of man 
or animals) show marked differences with regard to their special 
habitat in the body of their host; the lesions they produce will then 
show a peculiar localisation. Closely allied species may further 
differ in their geographical distribution which is indicated by the 
geographical occurrence of the respective lesions. It is, therefore, 
a priori, imaginable that the localisation and the geographical 
occurrence of certain symptoms may in certain cases be a valuable 
support for the distinction of the species which cause them. A 
different question is whether variations observed in the localisation 
and the geographical distribution of certain symptoms may be used 
as proof that they are due to different species of parasites. In 
this connection I must point out that, quite generally speaking, 
observations of the alleged sort are, in principle, statistical. 
I do not underestimate the value which statistical observations may 
have under various circumstances; but it is a fact also that from the 
same statistics more or less opposite conclusions may be 
drawn according to the point of view from which they are looked at. 
I may add that the statistics themselves are by no means all of them 
equally reliable. On the whole, therefore, I think that it will always 
be wise to test statistical observations very carefully before con¬ 
sidering what they seem to show, as an objective proof of some theory. 
An excellent example of the truth of what has just been said will be 
discussed towards the end of this article. 

In the case which at present occupies us Dr. SAMBON uses 
statistical observations referring to the localisation and geographical 
distribution of certain lesions as additional proofs of the existence of 
a species of parasite which is zoologically utterly doubtful. Judging 
a priori, I would consider the species as established, notwithstanding, 
when the additional proofs were binding and did not leave any 



i63 

visible gap. I am sorry to say that, from my point of view, Dr. 
Sambon’s proofs do not answer this description. 

The second difference between Sch. hamatobium and Sch. 
mansotti is said to be given in the different anatomical habitat, and 
the specific pathogenic action of the latter form. We will start with 
the well-established clinical fact that terminal-spined ova alone are 
voided from the bladder, whereas, lateral-spined are met with in the 
faeces. The conclusion generally drawn from this observation, and 
also brought forward in the discussion above mentioned by Sir 
PATRICK Manson, is that the lesions of the bladder are 
caused by worms producing terminal-spined ova, whilst the 
almost identical lesions of the rectum are caused by worms 
producing lateral-spined ova ; in other words, that the last-named ova 
appearing in the faeces are derived from the rectal 
lesions. In connection with this point, Sir PATRICK MANSON 
asked whether anybody had ever seen a lateral-spined egg in the 
urine. Nobody could answer in the affirmative; indeed, I do not 
remember myself to have specially noticed a lateral-spined egg in 
the urine. Thus far, observations agree very well; however, what 
I should like to point out is that even if I, or anyone else, had seen 
a lateral : spined egg in the urine, I would be unable to consider this as 
a fact of fundamental importance. To me it would appear as an 
accidental exception, due to accidental reasons, to the rule that the 
urine contains terminal-spined eggs only. One may examine the 
faeces of a thousand people without finding an apple-maggot, and in 
the faeces of the thousand and first there is one; the former observa¬ 
tions show that maggots are not a normal appearance in human 
faeces; the latter observation does not at once demonstrate the con¬ 
trary, but only shows that it is an occasional exception, the chief 
interest of which would lie in the question as to the conditions under 
which the exception occurs. As I have said, I would look at it from 
this point of view, should a lateral-spined egg some time be found in 
the urine. 

Passing to the supposed causal connection of the lateral-spined 
ova with the rectal lesions, I must confess that up to a few years past 
I shared the opinion that the former were derived from the latter. I 
must state to-day that this was a mistake; in many cases the 
lateral-spined eggs do not come from the 

M 



164 


rectal lesions. Desiring, some years ago, to make a 
drawing of a lateral-spined ovum, and having no faecal material at 
my disposal I took a papilloma of a preserved rectum and examined 
for ova. There were plenty of terminal-spined, but not a single 
lateral-spined could be discovered. New preparations made from 
other papillomata of the same rectum, gave no better results. Serial 
sections made of other recta showed similar conditions, in so far as 
sometimes terminal-spined eggs alone were found, sometimes both 
forms mixed, In no case, however, have I so far found, in the rectal 
wall, the lateral-spined eggs quite alone. The point most important 
in the present connection is that these observations leave no doubt 
that the vesical and rectal lesions so similar to each other in external 
appearance also contain one form of egg only. 
After the theory of Dr. SAMBON, this form is distinctive of Sch. 
hcematobium. Sch. hcematobium is, therefore, capable of producing 
rectal as well as vesical lesions; I see no reason w'hy, under 
peculiar circumstances, it should not be able to produce, 
in one case, vesical lesions alone, and, in another case, 
rectal lesions alone. The question as to what these 
peculiar circumstances may be is certainly of great interest, 
but it is secondary to the fact that both forms of lesions may be 
produced by Sch. hcematobium. ‘ Sch. mansoni' is said by Dr. 
SAMBON to have ‘ a specific pathogenic action 9 ; since Sch. hcemato- 
bium may produce identical lesions the alleged specificity does not 
exist, or shows, at least, a very remarkable flaw. 

We now come to a more important point. If lateral-spined ova 
do not occur at all, or occur in insignificant numbers only, in 
the rectal lesions it is impossible that such ova when they appear in 
the faeces can be derived from the rectal lesions. I had, in a 
number of cases selected entirely at random, found no trace of 
lateral-spined ova in the rectal papillomata ; where, then, could the 
lateral-spined ova come from? Before advancing further I must 
mention several facts which, in addition to the occurrence of both 
shapes of eggs in one and the same individual, figure as arguments 
in my theory as to the nature and the significance of the lateral- 
spined eggs. 

The place where the Bilharzia worms are, in post mortems, usually 
looked for, and most easily found, is the portal system. I have, since 



1893, seen a good many of them ; a fact which struck me from the 
beginning was their very different size. In certain cases, they 
presented about their normal dimensions; in others, they were 
markedly smaller, and in some, they hardly reached a third of their 
normal length. Another fact which sometimes very forcibly 
obtruded itself to the eye was that the specimens present in an 
individual case were, among themselves, of very much the 
same size, i.e., of about the same age. I still 
possess in my collection the material from one case, which consists, 
after specimens have been given away, and others have been used for 
examination, of 62 males, all varying in length from 3 to 4 mm. 
according to their somewhat different state of contraction. There 
are, in addition, females (though in fragments only) which must have 
measured from 5 to 6 mm. so far as their length is still determinable. 
I also remember another case in which the worms—m ales alone 
—presented two different sizes so distinctly that it was not difficult 
to separate them into two lots, each, of specimens about equal in size. 
On microscopical examination, all specimens proved to be sexually 
immature, and the degree of sexual development coincided about 
with their size. In many cases males were present alone; 
where both males and females were found they were still isolated; 
only in some two or three cases could a coupled pair be detected in 
the portal veins. The more advanced females contained one or a 
few ova in their uteri, all of them of the lateral- spined type, 
some of quite unusual shape. These observations only confirmed 
what had been seen and described by some former writers. 

As a helminthologist I have not limited my investigations to the 
parasites of man, but have carefully and through many years, studied 
—anatomically and biologically—the Trematodes parasitic in animals. 
One result of these studies was that, very generally, Trematodes at the 
approach of their sexual maturity were found to form abnormally 
shaped eggs. In some most interesting instances the female 
genital apparatus was, owing to some malformation, found completely 
shut off from the male apparatus ; there was no possibility for the 
egg-cells to become fertilized, but, nevertheless, the uterus was filled 
(in one case packed) with ova, all misshapen. In younger 
but normal specimens of the same species, the uterus contained more 
or less numerous normal eggs, but in front, there were, sometimes a 



166 


few, in other cases more, and in others again, crowds of the same 
abnormal eggs as had been seen in the specimens with the internal 
defects. In order to fully understand these statements one must, of 
course, have some knowledge of the anatomy and biology of the 
parasitic worms in general. I do not expect the ordinary medical 
man to have them, nor does he want them ; but I strongly recommend 
studies of the sort to all those who indulge in 4 formulating ideas * 
with reference to helminthological questions. Anyone 
would be laughed at if he tried to write a tale in a language of which 
he did not know the alphabet; but I might quote dozens of passages 
from modern papers on helminthological subjects which leave no 
doubt that the author did not know the significance of the terms he 
used 

Putting the facts observed in various species of Trematodes 
together with what had been seen by some earlier observers and 
myself in the young Bilharzia worms, I came to the conclusion that 
the lateral-spined must be abnormal eggs. I added that 
unimpregnated or isolated females would, perhaps, be ‘ unable to 
produce other than such abnormal eggs.’ I do not claim that this 
interpretation is the correct one ; but I daresay that it is based on 
a series of actual facts observed in the nearest natural relatives of the 
Bilharzia worms; in other words, that it is a quite well-founded 
4 conclusion from analogy.* 

Dr. SAMBON, speaking of this theory of mine, refers to an obser¬ 
vation of Dr. LEIPER, in which a terminal-spined egg was seen in an 
4 immature * female, and concludes that by this observation my theory 
4 is disposed of.’ I cannot help finding that Dr. SAMBON is some¬ 
what hasty in disposing of theories which are in contradiction with 
his own. I see that my young friend LEIPER states the immature 
condition of the specimen, but I do not see that he states the absence 
of spermatozoa in her oviduct. Was the worm, therefore, fertilized, 
or was it not? I further think that for everyone who will look at 
the case with an open mind it is clear that there is no mathematical 
line of demarcation between 4 maturity * and 4 immaturity.* The 
eggs are formed in the 4 ootype ’ which is situated at about the 
middle of the body at the posterior end of the long uterus. In the 
young females found in the portal vein the eggs are lateral-spined. 
They are gradually pushed along the uterus till at the end they are 



167 


expelled by the genital aperture situated behind the ventral sucker. 
Other eggs may follow the first, but, according to our present know¬ 
ledge, the number of those present at a time remains limited to 5 or 6. 
When the female is impregnated the formation of normal eggs begins. 
At about this period we ought to expect the uterus to show, in its 
hindmost part, a number of terminal-spined eggs, while the anterior part 
may still contain one or some lateral-spined ova. Such was the 
case in the specimen observed by BlLHARZ; the analogy it presents 
to some of the Trematodes described above is complete. Had 
BlLHARZ happened to see the specimen an hour or a day later the 
last lateral-spined egg would have been laid ; the specimen would 
have been 1 mature.* Had he happened to see it a day or a week 
earlier, no terminal-spined eggs might have been formed yet; the 
specimen was ‘ immature.’ As he actually saw it it was half 
‘ mature * and half ‘ immature.’ For the moment, therefore, I see no 
reason why my theory should be annihilated by the one accidental 
observation of Dr. LEIPER,—admitting even that the egg in question 
were really terminal-spined, and did not only appear as such because 
the lateral spine was turned towards, or away from, the observer. 

Dr. LEIPER himself says in the discussion (p. 45) that his obser¬ 
vation makes him believe * that the explanation was not correct which 
relied solely upon immaturity as the cause of the lateral spine.’ This 
is quite right, but I have not pretended either, that immaturity is the 
sole cause of the lateral spin£ Dr. SAMBON, in quoting my theory, 
makes me say that ‘ the eggs bearing a terminal (obviously a misprint 
for “ lateral ”) spine probably represent the product of unfertilized 
females.’ He thus does not notice that there is a slight but very 
important difference between saying ‘lateral-spined eggs are the 
product of unfertilized females,’ and saying, as I really have done, 
that ‘unfertilized females are not capable of producing other than 
abnormal eggs.’ As a matter of fact, several earlier authors have 
pointed out how fertilized females might, under certain conditions, 
produce lateral-spined eggs also. These suppositions have up to the 
present day not been proved as true, but they have not been disproved 
either, and it is at least not impossible that what those authors 
surmise may actually happen. At any rate, I have never pretended, 
and do not pretend, that immaturity is the sole cause of the lateral 
spine; nor is it impossible that immature females, although producing 



as a rule, lateral-spined eggs, may not, as an exception, produce one 
or another terminal-spined egg. Biological processes can never be 
pressed into a mathematical formula to which there is no possible 
exception. 

Speaking of the significance of these eggs I will provisionally 
quote the opinion of HOLCOMB, who says (1907, p. 62): 1 The West 
Indian infection proves that the lateral-spined eggs are not the eggs 
of unfertilized females, and some of my cases, which were under 
observation for one year or more, show only too well the persistence 
of the type of egg cannot be attributed only to young females.’ 
Before I can respond to this argument several other points must be 
discussed: I, therefore, at this place, limit myself to quoting 
HOLCOMB’S objection, and will return to the point later. 

The habitat of the mature Bilharzia worms are the finer 
ramifications, in the first place, of the vesical and, in the second place, 
of the rectal veins. As a logical consequence of my theory, one 
ought to expect that, there, they produce terminal-spined eggs 
only. Observation shows that the lesions actually contain such eggs 
in enormous numbers, and very often absolutely alone. 
Some stray lateral-spined eggs found at the one place or the other 
would not shake this rule. Even when large numbers were found 
in numerous cases the fact would not prove any specific nature of 
these eggs unless it were shown, either that unimpregnated females 
cannot possibly get to the same places, or, that impregnated females 
cannot under any circumstances form lateral-spined eggs. 

The young females living in the portal system produce lateral- 
spined ova, and successively expel them into the surrounding blood. 
Since the ova are not by themselves mobile the blood stream will 
carry them deeper into the liver, where, logically, they must finally 
become arrested in those vessels whose diameter equals their own. 
The theory held by me thus leads to the logical consequence that 
lateral-spined eggs must first and foremost accumulate in the liver. 
Their frequent and often plentiful occurrence in that organ is a well- 
known fact; indeed it represents another of the pillars on which my 
theory rests. According to the general belief (which I share) the 
worms grown up in the portal system reach their definite habitat in 
the pelvic organs by active wanderings, the vigorous males carrying 
the weaker females with them in the gynaecophoric canal. It is, 



i6g 

however, possible that the females are capable also of undertaking 
the wandering alone. As a matter of fact, isolated females have 
been seen in various veins ; but it is not sure whether they got there 
alone or by the help of males whom they afterwards abandoned. 
During this journey a 11 females go on laying eggs—at first 
abnormal ones, later (i.e., after they have become impregnated), 
normal ones. In all wider vessels, these eggs also are taken up 
by the blood stream and carried back to the liver where they join 
those which have arrived previously. There is, however, the probability 
that, now, terminal-spined ones may be among them; observation 
tells us that indeed these occur in the liver, though in numbers 
which vary considerably in the individual cases. But from what 
has been said above we may derive as the general rule that 
the lateral- spined eggs will prevail, the longer the females 
had to wait for fertilization, whereas the terminal-spined 
eggs will prevail, the sooner the females became fertilized. 

In the walls of bladder and rectum the worms make their way 
into the finer ramifications the diameter of which gradually becomes 
equal or even less than that of the male. From this point onwards 
it is difficult for the eggs laid by the female to escape into the general 
circulation. Pictures I have seen in sections of the vesical 
and rectal wall even seem to indicate that the females can stretch 
their (already thin) bodies to such an extent, escaping at the same 
time more or less from the gynaecophoric canal of the male, that 
their heads (close to which the genital aperture is situated) reach 
very fine capillaries. Eggs deposited there—either singly or in 
groups—would be kept in place by the walls of the vessels closing in 
upon them as soon as the female withdraws to her original place. 
The process may be repeated more or less often, a whole area 
becoming thus stuffed with ova. I have not seen the process 
here described actually going on; it is also probable that 
many variations occur; but the chief details are based on observed 
facts. 

The eggs, though originally deposited in the blood vessels, finally 
appear in the urine or the faeces: they must have passed through the 
tissues of the organs. I do not consider it as illogical to admit that 
what happens to the eggs in the walls of bladder and rectum may 
also happen to the eggs in the liver. Observation actually shows 



I/O 


them in the tissue of the organ: they, therefore, have left 
the blood vessels as they have in the pelvic organs. Admitting that 
they change their place in the tissue one will easily see that several 
things may happen. I will at this place only mention the possibility 
of some eggs getting into a blood vessel of the hepatic 
system. Should this take place the blood stream would carry 
them away from the liver; the next place where they are likely to 
become arrested again is the lung. Observation has shown that 
the organ in which, next to the liver, lateral-spined eggs are most 
commonly found i s the lung. What happened in the liver may 
happen in the lung; the eggs escaping from the latter would be 
carried by the arterial circulation to every possible organ. As a 
matter of fact, stray lateral-spined eggs have been seen at very 
different places. I will add that the way just described is not the 
only one by which they may reach the lung and other organs; 
however, these details may here be omitted as having no direct 
bearing on the questions under discussion. 

Returning to the eggs in the liver, there is, in addition to the 
possibility above mentioned, the other possibility of their getting into 
a part of the biliary system. In this case they would be 
carried to the gall bladder and thence into the intestine, from which 
they would be voided with the faeces. After 
I had ascertained that in certain cases lateral-spined eggs could not 
possibly be derived from rectal lesions, I came to think of this 
possibility as an explanation of their presence in the faeces. Obser¬ 
vation showed that the theory held good in this case also. In the 
first body available the first preparation made of the bile from the 
bladder revealed under the microscope four lateral-spined eggs; 
others were found in scrapings of the bladder wall, in the bile duct 
and all along the intestine. Three other cases examined subsequently 
presented similar conditions; I have not deemed it necessary to 
examine more. The theory had led to a conclusion which when 
tested by observation proved to be correct. 

This is one explanation I have to offer for the occurrence of the 
lateral-spined eggs in the faeces. There are others still, but I will 
not allow myself to enter upon details which have no direct bearing 
upon the question which here interests us. Observation has thus 
shown that the occurrence of lateral-spined eggs in the faeces of 



I?I 


living patients is not by any means a proof of the infection of the 
intestine proper, and quite especially of the rectum. These eggs 
may, and in many cases do, come from the liver; the only question 
of importance which remains to be answered in this connection is the 
question as to the reasons, why the infection of the liver is, 
in certain localities as the exception, in other localities as the 
rule, not followed by an infection of the bladder. I will show later 
that there is a possibility—and to my mind not even a very far¬ 
fetched possibility—to explain this curious difference, without the 
help of a mythical ‘ new species.’ 

We have seen above that the ‘ specific pathogenic action ’ assigned 
by Dr. SAMBON to his Sch. mansoni does not exist, for Sch. hcemato- 
biutn is capable of producing the same lesions. We see now that 
there is no * special anatomical habitat ’ either, for the lateral-spined 
eggs appearing in the faeces of living persons may be such of young 
Sch. hcematobium deposited in, and voided from, the liver. It may be 
added that up to the present nobody appears to have seen lateral- 
spined eggs in females imbedded in the gynaecophoric canal of the 
male, and the latter imbedded in a vein of the rectal wall. I have 
myself seen in situ quite a number of such females, but they only 
contained terminal-spined ova. I do not attribute any demonstrative 
value to these statements, but may point out that Dr. SAMBON’s 
theory would find an important support if he, or somebody else, could 
produce females collected under the conditions above 
mentioned, which possessed in her uterus exclusively, 
and as many lateral-spined eggs as the ordinary females possess 
terminal-spined eggs under the same conditions. I have in some 
instances counted the eggs in females collected from the mesenterial, 
rectal, and vesical veins, and have found them to vary in number 
between 80 and 150.* 

We now come to Dr. SAMBON’s third proof, the 4 peculiar 
geographical distribution’ of Sch. mansoni. I may freely confess 

* In a case quite recently examined I found a little colony of worms in the 
haemorrhoidal vein, about 7 cm. distant from the anus. There were five couples and 
two bachelor males. All males measure (after preservation) 5-6 mm. in length ; their 
testicles do not yet contain free spermatozoa. The females average 7 mm. in length. 
Their internal genital organs do not show spermatozoa. Three are also entirely free 
from eggs ; the two others contain each one lateral-spined ovum in the ootype, none 
in the uterus. This observation shows that the worms may leave the liver before 
sexual maturity is attained, but otherwise agrees with the theory. (Note added while 
reading the proofs.) 



172 


that when I first read the author’s own statements the statistical 
observations indeed seemed to strongly favour his view. However, 
on testing the evidence somewhat more seriously, I find that matters 
change their aspect considerably. Unfortunately, a number of the 
papers on which the evidence is based are not actually within my 
reach, and cannot, therefore, be compared. From those I possess 
I see that Dr. SAMBON quotes the literature in a rather unusual 
manner. 

Beginning with the West Indies it is true that according to the 
report by Dr. HOLCOMB (1907), rectal Bilharziosis is very common in 
those parts and vesical infection is rare. That the latter is actually 
absent cannot be said, for HOLCOMB enumerates four cases (one 
in a man from Guatemala, two in persons from Panama and one in a 
Porto Rican) in which the urine contained the terminal-spined 
ova, in one case even combined with the presence of lateral-spined 
ova in the faeces. Dr. SAMBON does not mention these cases, but 
only says that ‘ endemic haematuria ’ is not known in the West Indies. 
HOLCOMB states that he was not informed where, in the four cases, 
the infection was obtained Since it should not have been difficult 
to find out whether the infected persons had been to Africa, one may 
I think reasonably assume that the infection was acquired in loco. 
At any rate, there is no proof that it was not of local origin. 
However, I will not place great weight on these cases, owing to the 
fact that the place of infection is, though fairly clear, yet not 
positively ascertained. I consider it as more important that Dr. 
HOLCOMB has recently himself observed a case of urinary bilharziosis 
(information by letter). The most important case is that con¬ 
tracted at Martinique and very carefully studied by LETULLE (1905). 
LETULLE did not yet know of Sch. mans on i and the specific patho¬ 
genic action attributed to it. But he emphatically states that he 
found the bladder entirely free from infection. In the intestinal 
lesions, the lateral-spined eggs of ' Sch. mansoni 9 were seen in 
company with the terminal-spined eggs of Sch . hcematobium. 
The dimensions of all of them, by the way, agree very well with those 
of Sch. hcematobium, if one remembers the fact that the latter increase 
considerably in size during their embryonic development. Dr. 
SAMBON mentions LETULLE’s case as one of ' MANSON’s Bilhar¬ 
ziosis ’ (p. 32), but he does not mention that in this case (it is, so 



173 


far as I am aware, the first case of 4 MANSON’s Bilharziosis ’ 
thoroughly studied from a pathological point of view) both forms 
of eggs were found. 

Passing now to Africa, Sch. hcematobium is, according to Dr. 
SAMBON, alone present in Cape Colony. He refers to Harley’s 
observations and quotes from this author’s article- 4 In all my own 
cases I can positively say that only one form of egg has existed, 
namely, that with a terminal spine. Variation in the size, length, 
and outline of the egg is often observable, but I have never seen any 
egg with even a tendency to the formation of a side spine. I even 
doubt whether this peculiar form exists in the Distomum hcematobium 
itself.’ I have unfortunately no access to Harley’s paper, but 
LEUCKHART also mentions it, and he says: 4 Restricted exclusively 
to the possibility of examining the urine of his patients HARLEY had 
no knowledge of the existence of the eggs with lateral spines, and, 
therefore, considered the worms as a species different from that of 
Egypt ’ (P- 5 ° 7 )- It is thus true that HARLEY observed terminal- 
spined eggs only, but, unless LEUCKART’s remark is incorrect (for 
which assumption there is not the slighest reason), simply because 
he had no occasion to examine faeces in which the lateral 
spines are found. Dr. SAMBON then refers to the observations of 
Brock, and quotes: ‘that BROCK and others stated that they had 
never encountered the egg with the lateral spine.’ But, here again, 
BROCK himself says (p. 6): 4 1 have only been able to study the ova 
as they appear in the urine of patients suffering from Bilharzia/ I 
will not ask how it is possible that Dr. SAMBON makes such mis¬ 
leading statements in an article which apparently claims to be taken 
seriously. For there is no doubt that the observations of HARLEY 
and BROCK are anything but demonstrative of the absence of 
lateral-spined eggs in South Africa, as Dr. SAMBON makes it appear 
by his quotations. As to the observations made in the Congo Free 
State, I am sorry that I have no access to the original article, and, 
therefore, cannot say how far its contents correspond to the summary 
given by SAMBON. In the discussion Dr. Low states that in 
Uganda he saw exclusively rectal cases, but often also terminal- 
spined eggs 4 in the rectum ’ (1907, p. 45). 

Looking at this geographical evidence, as it now appears, the 
observer will first be struck by the fact that it has entirely lost its 



174 


original neatness. The statements, mostly based on clinical obser¬ 
vations during the daily routine work, seem contradictory, and 
nowhere is there a sharp line of demarcation left. I, for one, cannot 
see any trace of a ‘ peculiar geographical distribution ’ of the two 
shapes of the egg which are said to be distinctive of the alleged two 
species—admitting even that the statistics are all equally reliable, i.e., 
made with special regard to the question at issue. But, be that as 
it may; there is certainly no doubt about the evidence supplied by 
the case of LETULLE. This was, according to Dr. SAMBON’s own 
words, a case of ‘ Manson’s Bilharziosis,’ and, so far as I can judge, 
one absolutely typical both as regards the origin (Martinique) and the 
clinical and pathological aspects. Nevertheless, a careful study of 
the lesions revealed the presence in them of both forms of eggs. 
After SAMBON, it would thus have been a 1 combined * infection with 
Sch. mansoni and Sch. hcematobium ; an infection, however, in which 
Sch. hcematobium did not produce its own lesions, but those of 
'Sch. mansoni! It remains for Dr. SAMBON to show the way out 
of this labyrinth. For me there is no difficulty, for I say that both 
forms of eggs belong to the same species, and that the apparent 
differences between vesical and rectal Bilharziosis are not due to a 
difference in the species of the parasite, but to reasons which must 
be looked for elsewhere; we shall see later what they may be. If 
I were to make a * prophecy ’ I would say that in almost all cases 
of ‘ MANSON’s Bilharziosis,’ if they are so thoroughly examined as the 
Martinique case was by LETULLE, the eggs of Sch. hcematobium 
will be found among the eggs of ‘ Sch. mansoni! (I have just 
emphasized the word 4 almost ’; we shall see later that there are 
certain conditions under which the lateral-spined ought to be present 
quite alone). I especially recommend for examination the liver. 
It is a pity that it was not studied in LETULLE’s case; but I can 
easily comprehend that there was for LETULLE no visible reason to 
look for ova there. 

If we now compare the various pictures offered by Bilharziosis 
according to observers with those known from Egypt, there is 
no difference left except one, and this, as I must frankly confess, is a 
very striking one,-namely, the apparent irregularity in 
the localisation of the lesions. In order to make 
my case complete I will try to show that the biology, such as I 



*75 

interpret it, of Sch. hcematobiuni is perfectly sufficient to throw light 
on this difference also. 

Before advancing any further, and in order to avoid any misunder¬ 
standings, I will repeat that I do not ignore that what I have said 
with regard to the nature of the lateral-spined eggs, and what I am 
going to say hereafter with regard to the differences in the clinical 
aspect of the disease, is a ‘ theory/ inasmuch as it has not yet been 
established by experimental proof. In the absence of such proof, 
the only thing the scientific man, desirous of advancing our know¬ 
ledge, can do, is to collect carefully as many isolated facts as may be 
obtainable; to separate those which are (presumably) essential from 
those which are (presumably) accidental, and to piece all of them 
together into a continuous train of events. This is what I have 
been endeavouring to do ; I cannot imagine that a theory thus built up 
can be wrong in its fundamentals. It must, of course, be incomplete, 
or may be erroneous in details. I have already pointed out that 
Bilharziosis, in its varying aspects, presents a peculiarly complex 
problem, both as regards its pathogeny and the biology of the 
parasite. I do not think that I am wrong when I say that the latter 
represents the basis of the former, especially so far as the develop- 
ment and the behaviour of the worm within the human body are 
concerned. When, with regard to this part, I have knowledge of a 
good number of details, I owe that to the kind collaboration of my 
colleagues of the Medical School and the Kasr el Aini Hospital, Dr. 
ELLIOT Smith, the Anatomist, Drs. Symmers (now of Queen’s 
College, Belfast) and FERGUSON, Pathologists, Dr. Madden and Mr. 
Fr. Milton, Surgeons, who have discussed with me the observa¬ 
tions they had occasion to make during their professional work, and 
have given me many a valuable hint as to details with which I am less 
familiar. A priori, the various observations might have been ex¬ 
plained in various ways, but the right explanation could only be one 
which fitted in with the biology of the parasite. Sch. hcematobiuni has 
thus far successfully resisted all attempts at revealing the secret of 
its development. Nevertheless, we know a number of facts which 
definitely settle certain details; as to others, all we can do at present 
is to accept what seems to be most probable. For me, everything 
is probable as soon as it has been demonstrated in the nearest natural 
relatives of the Sch. hcematobiuni , i.e., either in other Schistosomes, or 



176 


in the digenetic Trematodes. I have acted according to this 
principle in formulating my theory. As a matter of course, in doing 
this, I depend upon our present knowledge. It often happens that 
a theory which seems probable and natural at one time is at another 
upset by new facts which, though not -annihilating the older facts, 
yet make them appear in a different light. I cannot foretell at 
present what facts may be in store for us with regard to Bilharziosis, 
and, therefore, cannot say that the theory which I defend at present 
is the right one in every detail. But I think that I can claim 
that it is based upon a large number of anatomo-pathological and 
helminthological facts deliberately weighed and compared. I have 
thought these remarks necessary in the face of Dr. SAMBON’s allusion 
that myself, and all the other workers in Egypt, have not been able 
within long years to find the solution of a problem which according 
to him was easy enough after all 

So as to be quite impersonal I will myself draw attention to an 
important biological point which I am not yet able to sufficiently 
account for. I am convinced that the lateral-spined eggs are 
abnormal and, probably, unfertilized. Nevertheless, when they appear 
in the faeces they very often contain a fully-developed miracidium. 
If we suppose, with some earlier authors, that mature females are, 
under certain circumstances, still capable of producing such 
lateral-spined eggs, the dilemma would resolve itself into the 
question as to what these conditions are (spontaneous con¬ 
traction ; pressure of the surrounding organs, either accidental or 
owing to their movements ; &c.). I have already said that this point 
is not yet determined. To me it seems, on the whole, very 
little probable, that fully mature females continue to produce 
lateral-spined eggs. If this be true, the presence of the miracidia 
would forcibly indicate that the eggs are capable of developing by 
parthenogenesis. From what has been observed in the hermaphro¬ 
ditic Trematodes it appears that the eggs must be fertilized, in 
order to develop. A priori, one ought to expect the same also in the 
Trematodes with separate sexes. However, considering the unmis¬ 
takable disadvantages connected with this separation in regard to 
the preservation of the race under the peculiar circumstances under 
which the Schistosomes are living, and considering further the very 
complicated development of many Trematodes, in which often several 



177 


asexual generations occur before the sexual stage is again reached, 
I would hesitate to pronounce a hasty conclusion. At any rate, the 
presence of fully developed miracidia in lateral-spined eggs is a point 
which still requires to be cleared up biologically. For the question 
as to the specific nature of the lateral-spined ovum the point is of no 
consequence; for its combined presence in the same individual with 
the terminal-spined egg is evidence enough that only individual 
conditions can be responsible for its formation. 

I will now try to show that the strange and striking difference 
offered by the clinical and pathological pictures of Bilharziosis in 
various places is not incapable of explanation if we consider the 
presumptive life history of the parasite, in connection with 
the habits of the host and the conditions 
of the country. In order to make this clear I must start 
from the beginning. 

The miracidium (often inappropriately called ‘ embryo ’) contains 
in its abdominal cavity the so-called 1 germinal cells/ the significance 
and ultimate fate of which are well known from their comparative 
study in various other Trematodes. The existence of these cells in 
the Bilharzia miracidium is absolute evidence that the 
miracidium cannot develop directly into an 
adult worm, but must pass through the stage of the ‘ sporo- 
cyst * which, in its turn, produces, either (and probably) at once, or 
by one or more intermediate generations, the definite worms. All 
attempts made by former authors to discover an intermediary host 
in which this development is gone through, have failed, and so have 
my own efforts. I have examined hundreds of specimens of all 
the molluscs common in the Nile valley, without finding any sporocyst 
which might have been brought into relation with the Bilharzia 
worms. I have placed quantities of free swimming miracidia in 
contact with the same molluscs, without obtaining an infection. It 
is easy to infect molluscs with miracidia of species which actually 
develop in them. I will not enter into details, but only say that the 
Bilharzia miracidia were never seen to take any notice of any mollusc 
in their neighbourhood, whereas others developing in a certain 
mollusc soon begin to swarm about it, and may, under the microscope, 
even be observed to enter into it. The same negative results were 
obtained with larvae of insects, with fishes, and with plants. I am 



178 

thus forced to the conviction that Man himself acts as 
intermediary host. 

If this conclusion is correct it leads to the important consequence 
that the spread of the Sch. hcematobium is not 
limited by the natural geographical distri¬ 
bution of a special intermediary host. It can 
spread wherever man carries it, so long as, and in so far as, the 
climatic and hydrographic conditions are favourable for its develop¬ 
ment. With regard to this point, I entirely disagree with Sir 
PATRICK MaNSON who says (1907, p. 653) that the peculiar geogra¬ 
phical limitations of Sch. hcematobium are difficult to explain if it 
does not require the services of an intermediary host. However, I 
also hold that Sch. hcematobium is by no means geographically so 
limited as it appears to be to the defenders of the existence of Sch. 
mansoni . 

No investigator has hitherto succeeded in keeping the miracidia 
alive for more than 30-40 hours; in my personal experiments, the 
upper limit found was 28 hours. They must find some new shelter 
within this time. If they are destined to return into man directly, two 
possibilities are, a priori, imaginable, viz., that they enter by the 
mouth, or that they enter by the skin. I have found by experiment 
that hydrochloric acid diluted with water to the extent of 
I : 2000 kills them within 2-3 minutes, a solution of I : 1000 almost 
instantaneously; by exclusion I am thus led to the view that they 
enter by the skin. There are some other facts which may be 
interpreted in favour of this view; but I will not mention them here. 
In Man, the miracidium must develop into a sporocyst which, either 
directly, or indirectly, generates the Bilharzia worms. 

We have already seen that the only organ of the body thus 
far known to harbour young, and sometimes very 
young, worms is the liver. I therefrom conclude that the liver is the 
habitat of the sporocyst, from which the worms later escape into the 
portal vein. A priori, one might think of the possibility that they 
can escape also into the hepatic veins. As a matter of fact, they 
have been found comparatively often in the vena cava (KARTULIS), 
the lung (Symmers), &c. If the liver is the seat of the sporocyst it 
is a curious coincidence (perhaps it is not a mere coincidence) that, 
in the known intermediary hosts of other Trematodes, it is the 
liver which harbours the sporocysts. 



I 79 


At post mortems, it is not uncommon to find males alone 
in the portal vein. These males are often conspicuously of the 
same size, in other words, all of the same age. They must have been 
generated at about the same time ; this would become comprehensible 
on the assumption that they are generated in one sporocyst. If 
one sporocyst may produce males I see no reason which forbids the 
assumption that the females take their origin in separate sporocysts. 
As females are, as a rule, found much more rarely than males, it may 
be admitted that male sporocysts are commoner than female. 

This is the way in which, according to the facts at present 
available I am forced to explain the arrival of the parasites in 
the human body. I will now describe how I seem to see the con¬ 
nection between the special aspects of the disease and the habits of 
the population as they are observable, in the first place, in Egypt. 
In Egypt, Bilharziosis is very common. In the towns it is especially 
the children who are infected; among our students, there are always 
some who have, or have had, haematuria. Some of them assert 
emphatically that they have got it while in the country. In all of 
them the disease lasts for some years and then disappears. All 
severe cases come from the country. The Egyptian peasants 
usually work their fields in companies; sometimes of two or three, 
sometimes of several dozens ; standing with their feet, and working 
with their hands, in the water or the mud. They often also bathe 
in companies in canals with slowly flowing water, pools, &c. One 
of them who is infected with urinary Bilharziosis, when urinating into 
the water, infects it with several hundreds, perhaps thousands, of 
eggs. In warm weather the miracidia hatch within a few minutes. 
They have at once the opportunity of finding a new shelter, either 
in the skin of the man who voided the eggs or in the legs or hands 
of one of his comrades working close by him. Many of the 
miracidia which enter the skin will not succeed in finding their way 
to the liver, but a few do so. These possibilities of infection are 
repeated every time a man urinates into the water. They are 
perhaps repeated every day the season of the Nile flood lasts. There 
is thus not only the possibility, but the extreme probability, that 
several miracidia attain their destination at short intervals. 

The worms they give rise to in the liver are of about the same 
age. On this supposition, viz., that several miracidia succeed in 
N 



i8o 


gaining the liver at short intervals, it becomes probable that, from 
the beginning, there will be males and females. In this case, 
the females, grown up to the sexual stage will not have to wait long 
for fertilization. They will produce a few abnormal eggs, but are 
soon taken up by the males and carried to the pelvic organs. On the 
whole, therefore, only a comparatively restricted number of lateral- 
spined eggs will be deposited in the liver; they may, subsequently, 
be joined by larger numbers of terminal-spined. 

The chief habitat of the adult worms is undoubtedly the bladder. 
The chief vein which leads them there is the inferior mesenteric vein. 

I will point out in passing, that during the journey an occasional 
couple, before reaching the vein named, may accidentally get into a 
side branch coming from some other part of the bowel. In such a 
case the worms would give rise to an isolated focus of infection, or 
a separate growth at an unusual place. Such unusually located 
lesions have often been observed, and are, I think, correctly explained 
in the way alluded to. The inferior mesenteric vein leads the worms 
to the haemorrhoidal plexus close to the anus, but not immediately to 
the bladder. In order to settle the very important anatomical point 
whether there is a connection, wide enough to let the worms pass, 
between the veins of the rectum and those of the bladder, Dr. ELLIOT 
SMITH has very kindly made a dissection of the respective parts. 
Since he proposes to return to this anatomical question in detail 
himself, I here limit myself to the statement that such connections 
exist, wide enough to allow, not one couple only, but two and perhaps 
even three to pass side by side. I have subsequently found the 
worms in veins which, to judge from their width and course, were such 
connecting branches between rectum and bladder. 

I will not omit in this connection to recall the remarkable degree 
in which Trematodes are able to contract their bodies. I have under 
the microscope followed Cercariae entering into tadpoles and insect 
larvae. The actual opening they make in the skin of these ‘ supple¬ 
mentary 9 hosts is often so small that the worms assume the shape of 
an hour glass; but they get through it, evidently without difficulty. 
There is, therefore, every probability that male Bilharzia worms may 
manage to travel through vessels the ordinary diameter of which does 
not exceed a half or even a third of a millimetre. 

There is thus no doubt that the parasites have a direct route from 



i8i 

the portal vein to the bladder. Another very important question is 
why they do not remain in the veins of the rectum (or the intestine in 
general, which is apparently the original habitat of the Schistosomes), 
and how they find their way through the (comparatively) few connec¬ 
tions between rectal and vesical veins into the latter. In order to 
explain this remarkable ‘ knowledge of anatomy * I will draw attention 
to some well-known facts derived from the comparative biology of 
other parasitic worms. The larvae of the Filaria bancrofti , e.g., 
after having been sucked up by a mosquito, leave the intestine by 
perforating its wall, and make their way into the thoracic muscles ; the 
larvae of the Filaria immitis do the same, but seek the Malpighian 
tubes. The mature Ankylostoma worms do not live irregularly 
scattered throughout the small intestine, but chiefly accumulate in a 
certain region. Many Amphistomes inhabit the first stomach of 
their hosts (Ruminants), but the specimens found there are, according 
to my personal experience, never below a certain size. The young 
stages live, often by hundreds, in the small intestine. They have 
been swallowed along with the food, but do not at once settle in the 
stomach (which they have to traverse in order to get to the 
small intestine); it is not until they have reached a certain size at 
this provisional habitat in the small intestine that they return to 
the first stomach which is their definite habitat. 

In all these cases the worms must be guided by something which 
makes them find their place of destination. I have no doubt that 
this something is given in the peculiar chemical composition of the 
organs, or the juices, at the respective places; in other words, the 
wanderings come under the phenomena of ‘ chemiotaxis.’ One 
might suppose that the conditions in the small intestine of man are 
about the same throughout its total length (at least behind the 
entrance of the bile ducts). But the fact that the Ankylostoma worms 
normally settle in the anterior half only, is to my mind evidence that 
there must be differences which to the worms are noticeable, and lead 
them to select one special place in preference to any other. The 
fact that stray specimens may often be found more or less far away 
from this place, does not shake the rule; these specimens are ‘ the 
exceptions which strengthen the rule.* 

Starting from these reflections I conclude that the Bilharzia worms, 
also, are guided in their journey by chemiotactic influences. I do 



182 


not think it unreasonable to conceive that the venous blood coming 
from the bladder is chemically slightly different from that of the 
rectal veins, and that this difference, slight as it may be, exercises an 
attractive influence on the worms, thus ‘ leading ’ them to the bladder. 
It is in this connection certainly not without significance that the 
whole journey goes against the blood stream, just as the dog 
scents the game against the wind, but not with the wind At any 
rate, the veins of the bladder seem to be those first sought by the 
worms, although the rectal veins are nearer and would serve their 
purpose (to bring the eggs to some place where they can easily reach 
the outer world) equally well. As a matter of fact, the other 
Schistosoma species known are chiefly inhabitants of the intestinal 
veins. In Sell, lucniatobium the first infection of previously healthy 
persons seems to normally concern the bladder, whereas (apparently) 
the rectum becomes implicated after repeated infections only. One 
might almost imagine that after numerous eggs have been deposited 
in the bladder, and the normal function of the organ has become more 
or less impaired, the blood loses for the worms its peculiar ‘ scent.’ 
There may also be mechanical reasons which keep them in the rectal 
veins in larger numbers than before, &c. In this, or some similar 
way, the rectum would gradually become infected after the bladder. 
However, I do not find any reasonable objection either, to the assump¬ 
tion that in some eases some couples of worms might from the 
beginning remain and establish themselves in the veins of the rectum. 
Owing to the kindness of Dr. Ferguson, 1 have recently had the 
opportunity of examining several cases of ‘ early Bilharziosis of the 
bladder,’ in which the most scrupulous inspection of the rectum could 
not detect any visible change in the normal aspect of that organ. 
Nevertheless, quite a number of (terminal-spined) eggs were found 
in the residue after a part of the rectal wall had been macerated in 
caustic potash. 

In bladder (or rectum) the real oviposition begins ; the eggs are 
at first scarce, but gradually increase in number. They are laid in 
the blood vessels but afterwards escape into the tissues and are 
finally voided from the body after having traversed the mucous 
membrane of the bladder (or rectum). We do not yet know how 
long it takes them to accomplish this journey, but some will reach the 
end of it in a comparatively short time, whereas others may not 



i83 


succeed even after a long time. In any case, they do not appear in 
the urine (or the faeces) at once. The beginning of the haematuria 
is quite insidious; from Egypt many cases are known where there 
was not even haematuria; the eggs were accidentally discovered in 
examinations of the urine made for other purposes. 

While travelling through the tissues the eggs undergo their 
embryonic development. The eggs at any time visible in the uterus 
of the female worms invariably contain an undivided egg-cell. As 
the embryo develops within the egg shell, the egg itself increases in 
size; that with a mature miracidium inside measures, on an average, 
o'13 to 0*15 by 0*04 to 0*06 mm, whereas immediately after formation 
in the ootype it is only o'o8 to o'09 by o’Oj to 0*04 mm. in dimension. 
During the embryonic development, many embryos die. Their 
bodies become gradually decomposed, and afterwards replaced by 
calcareous masses; the eggs become ‘ calcified.’ Their appearance 
in this state is known well enough, so I do not want to dwell on it. 
With the moment of the death of the embryo, the increase in size of 
the whole egg is stopped. The calcified eggs thus present the well- 
known variations in size; they remain small when they died early, 
they are larger when they died later. 

Appearances I have seen in many sections of bilharzial tissues (of 
recent and old cases) make me believe that dead eggs are no 
longer capable of traversing the tissues as 
easily as living ones do. They will thus remain in the 
tissues more easily. In cases of very long standing they are often 
found quite alone; in other words, cases in which calcified eggs are 
found in the tissues, or voided with the excreta, are old cases. 

The age the worms may reach is not yet known; for the sake of 
my argument we will assume that it be three years. Three years, 
therefore, after a man has become infected (and has had no occasion 
since to become re-infected!) adult worms will no longer be in his 
system. But during their three years’ life the females have produced 
an immense quantity of eggs. Many of them have been voided 
during the same time, but as many, probably many more, are still in 
the tissues, and continue to be voided with the urine (or the faeces) 
for a more or less longer period. But their presence, and even the 
presence of a live miracidium in them, is by no means a sign that the 
worms which produced the eggs are still alive. I may mention in 



this connection that practitioners have now and again tried to relieve 
their patients by administering drugs with the view of killing the adult 
worms. Considering what has just been said, and considering the 
other fact that the great majority of Bilharzia patients do not come 
for relief before the evil is more or less far advanced, it will at once 
be understood that in nine cases out of ten every attempt at the worms 
will be too late. That the eggs voided still harbour a live and 
active miracidiiim is no proof that they must be derived from a com¬ 
paratively recent infection. We know of cases which, according to 
our present knowledge, cannot be explained unless by the assumption 
that within the human body the miracidia enclosed in their egg shells 
are capable of retaining their vitality for many years. The fact is 
by no means an uncommon one among the parasitic worms; the 
encapsuled Trichina or the wheat-eel dried up in its grain are well- 
known instances of longevity, which also show its biological signi¬ 
ficance. For it is not difficult to understand that the longer an 
immature parasite is able to wait for its chance, the greater becomes 
the probability that it obtains the chance for getting under those 
conditions which allow it to grow to sexual maturity and 
propagate its race. I think that the capability of the miracidia, in 
the eggs which are not at once voided from the body, to remain alive 
for a very long time must be looked at from this same point of view. 

We have so far considered (theoretically) what I should like to 
call the normal course of events, i.e., that which takes place in 
localities where it is comparatively easy for the miracidia to find a 
new host wifhin a short time. Under these conditions there is every 
probability that the females have not long to wait for the males. They 
produce few abnormal ova, the liver remains almost free, but terminal- 
spined ova are deposited plentifully in the bladder or rectum, as the 
case may be. There is 4 urinary Bilharziosis ’ characterised by the 
apparition of terminal-spined eggs in the urine; the same eggs may 
also appear in the faeces, but lateral-spined ones will be so scarce 
that they seem to be altogether absent. We will now consider what 
is likely to happen under conditions which are no longer so 
favourable to the worms. 

A man, for instance, does not work, or bathe, in the water day by 
day, but only at intervals of weeks or months; he does not remain 
in the water long, but for some minutes or an hour only; he avoids 



l8 5 


the close company of others ; the water itself may perhaps be quickly 
flowing, thus sweeping the miracidia away from the place where they 
have been hatched, &c., &c. Under all these conditions, combined 
with the short time the miracidia are able to remain alive in water, 
the chances of entering the skin of a new human host are 
considerably reduced for any which may be in the water. Many a 
time not one will succeed in finding him and entering his body. On 
a single occasion, however, a few miracidia manage to enter his skin, 
and one gets safely to the liver. It produces males (we know that 
these are much commoner than the females; the probability of 
picking up a male sporocyst is therefore greater). The worms grow 
to sexual maturity, but finding no females they wait perhaps for a 
certain time, and then undertake the journey to the pelvic organs 
alone. After some time, the liver is again free from worms; the 
infection, although it has taken place, remains without consequences. 

The man continues exposing himself to the conditions for 
infection as indicated. What happened previously may be repeated 
at intervals, but on one of these occasions a miracidium may enter 
his body which produces female worms. These in due time begin to 
lay lateral-spined eggs. The oviposition goes on, perhaps, for a long 
time. The number of lateral-spined eggs increases steadily; all are 
carried to the liver. It is possible (I might say probable) that some of 
the females try to undertake their journey alone, but owing to their 
inferior muscular strength they may sooner or later be driven into 
some smaller side branch of the mesenterial, chiefly of the splenic 
and inferior mesenteric veins. Not one succeeds in making the entire 
long journey to the rectum and the bladder. Of the specimens that 
have left the chief track leading to these organs, one or the other may 
reach the wall of the large bowel, filling a small area with her lateral- 
spined eggs. At the end there will be a comparatively strong infection 
of the liver, and perhaps some isolated infected patches in the wall of 
the intestine, but no terminal-spined ova will ever appear, nor will 
there be a regular infection of the bladder. After some time the 
lateral-spined eggs of the liver begin to appear in the faeces, and 
they continue being voided in this way for several years. I have 
further above (p. 174) hinted at the possible existence of cases in 
which even the most careful post mortem examination would not 
detect any terminal-spined ovum in any organ; we here have the 



i86 


conditions under which such cases must arise. This seems to 
me a suitable place, too, for inserting a biological remark. 1 have 
spoken above of the presence of fully developed miricidia in lateral- 
spined eggs. If these eggs are unfertilized, as I hold they are, they 
must be capable of developing by parthenogenesis. We now see the 
vital advantage the parasites would derive from such a capability for 
the propagation of their race in localities where the conditions for 
infection are scarce. I may confess that from this point of view the 
hypothesis loses for me much of its original strangeness. 

I am now able to answer the objection raised against my 
interpretation of the lateral-spined eggs by HOLCOMB. I suppose 
that the author means to say in his argument that the lateral-spined 
eggs could not have been derived from young females because in the 
course of the year during which the eggs were observed the females, 
though young perhaps initially, ought to have grown to sexual 
maturity and thus passed on to the formation of terminal-spined 
eggs; in other words, during a year the eggs appearing in the faeces 
ought to have changed from the lateral-spined to the terminal-spined 
type. This objection of HOLCOMB would indeed hold good if the 
Bilharzia eggs, like those of the intestinal parasites, were voided from 
the body of the host within 24 or 48 hours after their oviposition. But 
we know that they come from the liver, in which they have been 
laid by young or unfertilized females within a comparatively short 
period, but from which they are voided as gradually as the terminal- 
spined eggs are from the bladder—quite irrespective of what has in 
the meantime become of the worms which laid them. The 
observation referred to by HOLCOMB is therefore no proof against 
my interpretation that the lateral-spined eggs are the products of 
young, or unfertilized, females. It is, on the contrary, an argument 
in favour of it, whereas it would have been a certain evidence 
against my views had HOLCOMB observed that the eggs changed 
their shape in a marked degree in so short a period. 

If we admit attempts at independent wanderings on the part of the 
females, it may happen that some of them, succeeding in getting near 
the haemorrhoidal plexus of the rectum, may find there some males, 
remnants of a previous infection. Or else the host may contract a 
new infection with another male sporocyst while some females of a 
previous infection are still alive in the liver. In both cases matters 



187 


would assume what we have called above their normal course. After 
due time terminal-spined ova would appear in the urine, while the 
lateral-spined eggs of the liver continue being voided by way of the 
rectum. I will not spin out this narrative any more. I think it will 
now be seen that in the way alluded to the clinical and pathological 
picture shown by the infection in any particular individual must 
depend upon the more or less favourable nature of the conditions of 
infection to which the individual has been exposed. A first infection 
with one or some male sporocysts would not lead to any consequence. 
A first infection with a female sporocyst would give a picture typical 
of ‘ MANSON s Bilharziosis/ i.e., an untouched bladder but lateral- 
spined eggs appearing for years in the faeces. In all countries where 
infection with Sch. haematobium is possible, a man once infected will, 
as a rule, be subject to the opportunity of re-infection. The aspect 
the disease will then show must vary with the intervals at which the 
infection becomes repeated, and with the sex of the worms which are 
acquired. We may get pictures such as represented by the case of 
LETULLE, where the external aspect still preserved the features of 
‘ Manson’s Bilharziosis/ but where, internally, the normal, terminal- 
spined eggs were found in company with the lateral-spined type. If, 
finally/a man once only in his life, and perhaps for a few hours only, 
happens to come under a peculiar combination of circumstances which 
favour a simultaneous entrance of a larger number of miricidia, even 
in a country where otherwise the conditions for infection are 
unfavourable, he will contract for perhaps three, perhaps six, perhaps 
more years 4 urinary Bilharziosis/ either pure, when he was not 
previously infected, or mixed with 4 MANSON’s Bilharziosis/ when he 
was infected with this peculiar type before. I am personally perfectly 
sure that the four cases of urinary Bilharziosis quoted by HOLCOMB 
were contracted in loco after the fashion here described. 

On the whole, therefore, I do not, from my point of view, see any 
sharp line of demarcation between the two types. They are simply 
the opposite ends of a continuous series of intermediary stages. 

I cannot quit this subject without drawing attention to another 
point which seems to me of great interest. We have seen that 
LETULLE expressly states that in his case the bladder was entirely 
free from infection. I can only interpret this statement in the sense 
that no pathological changes were perceivable in the bladder, but I 



188 


cannot quite believe that a close examination would not for all that 
have resulted in the detection of some eggs in the tissue of the 
bladder. It would have been of the utmost interest to know of what 
type they were. Cases like Letulle’s are rare in Egypt, because 
of the specially favourable conditions for infection in this country. 
But other observers may have an easier opportunity of examining 
them. Instead of a tedious preparation of sections, it would suffice 
to macerate a piece of the bladder-wall in caustic potash, and examine 
the residue. The shells of the Bilharzia eggs are not at once dissolved 
by this reagent, and even a few eggs would be found without 
difficulty, if present. 

The conditions unfavourable for infection as they were suggested 
above will in general obtain in countries where there is a thin 
population, where the people do not come regularly in contact with 
water every day for a long part of the year, where they are not in the 
habit of working (or bathing) in companies, where there is not much 
water, or where the water, though abundant, flows quickly, &c., &c. 
I know neither the country nor the habits of the population in South 
Africa, in Uganda, in the Congo Free State, or in the West Indies, 
but I am fairly sure that on close observation of details the special 
aspect of the disease will be found to vary in accordance with the 
conditions for infection as they have been specified above. 

I have previously alluded to the relative value of statistical 
observations. I can now illustrate what has been said there by an 
instructive example. HOLCOMB (who believes in 1 Sch. mansoni *) 
after having given an extensive account of the cases of intestinal 
Bilharziosis observed in tropical America, compares them with the 
statistics published by various observers on the relative frequency of 
the different forms of Egyptian Bilharziosis (1907, p. 59). The main 
result he comes to is that, because there is in Cairo on an average one 
case of intestinal to 17 cases of vesical Bilharziosis, one ought to 
expect the same for the West Indies, if the cause of the disease were 
the same parasite in both countries. The actual observations, 
however, show the contrary: the intestinal form is very common and 
the vesical form is extremely rare; therefore, it appears that in the 
West Indies there is also another species of parasite. 

For me, the same statistics do not prove anything beyond the 
bare fact that the disease shows in the two countries a remarkably 



different aspect. In order to find out the reason for this fact, I would 
deem it necessary to analyse, if possible, a 11 the factors that have, 
or may have, a share in bringing about the fact. The species of the 
parasite is one of them ; the species of the host is another. But 
there are others, and amongst these the local conditions for infection 
are to my mind a factor of the first practical importance. This factor 
has been completely overlooked by HOLCOMB when he drew his 
conclusions, although its value is, so far as I have a judgment, fully 
recognised in modern epidemiology. Let us only assume it were 
possible to take one of the West Indian Islands, make its climatical 
and hydrographical conditions absolutely like those of Egypt, make 
the population (which is slightly infected with intestinal Bilharziosis) 
as dense as it is in the Nile Valley, make it live and work after the 
fashion of the Egyptian fellah, and then shut the Island off entirely 
from communication with the rest of the world; I have little doubt 
what the statistics would say some ten or twenty years hence. 

Resuming now what has been said in this lengthy discussion, I 
must state that, of the evidence brought forward by Dr. SAMBON in 
order to justify the creation of ‘ Sch. mansoni *: i, the zoological 
proof is absolutely insufficient; 2, the anatomo-pathological proof 
does not stand any serious test; and 3, the geographical proof is 
based upon a peculiarly one-sided interpretation of the literature. In 
all the evidence there is not the slightest detail which would really 
point to the existence of a distinct species in the West Indies and 
certain parts of Africa. It would be unwise on my part to go so far 
as to contend that such a species, or perhaps even several species, 
can not, altogether, exist. This is quite possible from the zoological 
point of view ; but, zoologically, there is no possible doubt either that 
this species, or these species, must produce the same two 
shapes of eggs as does the Sch. hcematobium ,, or else our 
present information is wholly incorrect. If, therefore, Dr. Sambon 
wishes to maintain that there is an independent * Sch. mansoni 1 in the 
countries above-mentioned, the entire proof of its existence still 
remains to be given. 

I cannot conclude this article without making some remarks of 
remonstrance with regard to another passage in Dr. Sambon’s paper 
‘ On the Part played by Metazoan Parasites in Tropical Pathology/ 
Speaking of the infection with Agchylostoma duodenale , he says that 



igo 


the theory of infection by the skin 1 now stands on the firm foundation 
of experimental proof.’ But the printed abstract (Journ. Trop. Med. 
1908, p. 34) then goes on to say that ‘ Dr. SAMBON doubted, however, 
whether the trachea-oesophagus part of the journey was more than 
conjecture; he thought the larvae could certainly reach the intestine 
by a safer and more direct route.’ The author then refers to the 
larval stages of certain Cylicostomes found encysted in the intestinal 
wall, and to the larvae of Sclerostomum vulgare living in aneurysms 
of the mesenteric arteries, in horses. I presume that Dr. SAMBON 
means to indicate by this reference that there is a connection, or an 
analogy, between the development of the forms mentioned and that 
of the Ankylostomes. If my presumption is correct, I may say in 
answer that thoughts of this sort are as old as they are unfounded. 
They will be discussed in detail in the second part of my monograph 
on Ankylostoma which I am at present writing. If Dr. SAMBON 
further ‘ thinks ’ that the larvae could ‘ certainly ’ reach the intestine 
by a safer and more direct route, and if he ‘ doubts ’ whether the 
trachea-oesophagus part of the journey is ‘ more than conjecture,’ I 
cannot help it. I will only state the following facts. My actual 
observations concerning the wanderings of the larvae were first made 
known in a paper read before the Sixth International Zoological 
Congress at Berne in 1904 They are printed in the * Comptes 
rendus’ of this Congress (1905a, p. 225f.), and again described in 
connection with some other questions concerning Ankylostomiasis in 
a later paper of mine (1905b). In Berne I exhibited a series of 
microscopical sections showing the larvae in the different stages of 
their journey. These preparations afterwards went for some time 
to Dr. Oliver, of Newcastle-on-Tyne, who, with my authorisation, 
had lantern slides of some of them made which he used in connection 
with a paper read by him before the North of England Institute of 
Mining and Mechanical Engineers (1904). Some of these photographs 
were afterwards (I regret to say, without my authorisation) published 
in SIR Patrick Manson’s ‘ Lectures on Tropical Diseases’ (1905); 
there is one (on page 20) showing larvae in a bronchus, and another 
showing a larva in the larynx (on page 21 ; it is erroneously labelled : 

‘ in stomach ’). In 1906 I had the pleasure of presenting to the London 
School of Tropical Medicine a series of preparations, accompanied by 
a detailed description, of all the important stages of the journey of the 



larvae from the skin to the larynx. The tedious work of making 
these preparations was undertaken with the special purpose of 
sending them away in order to allow authors to form an individual 
opinion without great personal trouble, except, of course, that of 
looking at the preparations. Dr. SAMBON has not looked at them, 
nor has he consulted the literature before ‘ formulating his ideas.* I 
am sure that I do not under-estimate 4 The Importance of Rational 
Inductive Methods in advancing Knowledge ’ (Journ. Trop. Med. 
1908, p. 41), but I doubt whether f ideas ’ like these (and several others 
formulated by Dr. SAMBON with regard to helminthological 
questions) have a right to be classed under that heading. 

Cairo, 16/k March , 1908. 


REFERENCES 

Bilharz, Th. 1852. Ein Beitrag zur Helminthographia humana, in Zeitschr. wiss. 
Zool. IV, p. 53-76, Taf. 5. 

Brock, G. S. 1893. On the Bilharzia haematobia, in Journ. Path, and Bacteriol. 
II. Repr. 24 p. 3 PI. 

Harley, J. 1864. Endemic Haematuria of Cape of Good Hope, in Med.-Chir. 

Transact, p 55. (Not accessible, quoted after Leuckart, 1894, p. 506.) 
Holcomb, R. (\ 1907. The West Indian Bilharziosis in its Relation to the 

Schistosomum mansoni (Sambon, 1907), with Memoranda in ten Cases, in 
United States Xav. Med. Bull. I, Nr. 2, July, p. 55-80. 

Eku( kart, R. 1894. Die Parasiten dcs Menschen, etc. Leipzig 11ml Heidelberg, 
II. AufL I, p. 468-534. 

Looss, A. 1905a. Schistosomum japonicum Katsurada, eine neue asiatischc 
Bilharzia des Menschen, in Centrbl. Bact. I. Abth. Orig. XXXIX, p. 280-85. 
Looss, A. 1905b. Die Wanderung der Ancylostomum-und Strongvloides-Lai ven 
von der TTaut nach dem Darm, in C.R. 6mc Congr. Intern. Zool. Berne torq, 
n. 225-33. 

Looss, A. 1905c. Kinige Betrachtungen liber die Infection mit Ankylostonium 
duodenale von der Haut aus, in Zeitschr. klin. Med. LVIII. II. 1 and 2. 
Repr. 43 p. 

Manson, Sir P. 1905. Lectures on Tropical Diseases, London, Arch. Constable 
and Co. 

Manson, Sir P. 1907. Tropical Diseases. A Manual of the Diseases of Warm 
Climates. Fourth Ed. London, Cassell and Co. 

Sambon, L. ico7a. New or little known African Entozoa, in Journ. Trop. Med., 
X, Nr. 7, April 1st, p. 117. 

Sambon, L. 1007b. Remarks on Schistosomum mansoni, in Journ. Trop. Med., 
X, Nr. 18, Sept. 16th, p. 303-4. 

Sambon, L. 1908a. On the Part Played by Metazoan Parasites in Tropical 
Pathology, in Journ. Trop. Med. XI, Nr. 2, Jan. 15th, p. 29-36. 1908b. 

Discussion ibid, No. 3, Febr. 1st, p. 44-46. 




■93 


THE PREVENTION OF DENGUE FEVER 


BY 

E. H. ROSS, M.R.C.S.Eng., L.R.C.P.Lond. 

MEDICAL OFFICER OF HEALTH, PORT SAID; PUBLIC HEALTH INSPECTOR, SUEZ CANAL; 
EGYPTIAN PUBLIC HEALTH DEPARTMENT 


Received 25 March , 1908) 


Egypt has always been subject to periodical epidemics of dengue 
or dandy fever. In some of the towns the disease seems to be 
endemic, but sudden outbursts occur which spread all over the 
country. The disease presents the same characteristics as in other 
parts of the world and rarely gives rise to much difficulty in diagnosis. 
During epidemics the classical symptoms are very evident, including 
the pains, the apyretic period, and the rashes, which are sufficient to 
differentiate it from influenza. When pandemics of the disease occur 
in Egypt every town is invariably attacked, and few people escape. 
The death-rate, however, is very small as noticed elsewhere, though 
the debility and cardiac depression following an attack occasionally 
account for the sudden deaths of a few individuals who before were 
healthy. Since the discovery of the means of the transmission of 
malarial fevers it has been suggested by various writers that dengue 
fever is also conveyed from the sick to the healthy by the mosquito. 
Apparently Graham, of Beyrout, was the first to bring forward strong 
evidence of this, 1 and he named Culex fatigans Wied: as the 
culprit. Since that date further and conclusive evidence has been 
brought forward to support this statement. 2 

Dengue fever used to be very prevalent in Port Said, as in other 
parts of Egypt, up to the year 1905. An epidemic of the disease 
occurred in the town during the summer of 1904, and in the spring of 
1905. This epidemic was part of an infection of all the towns of 
Egypt, and was most severe. The hospitals were full of cases, and 
patients actually contracted the disease in them. In Port Said 
almost everyone suffered from an attack, and the place was regarded 



194 


as fever-stricken and unhealthy. The town was full of mosquitoes, 
including two species of Anophelines, Culex fatigans and Stegomyia 
spp., in abundance. These mosquitoes were breeding in cess-pools 
under the houses, in basement cellars flooded with sewage, garden 
fountains, barrels containing water, and were a veritable pest day and 
night, summer and winter. 

In May, 1906, a campaign against mosquitoes was instituted in 
the town as a general sanitary measure, with funds subscribed by the 
Egyptian Government and the Suez Canal Company, the support of 
Prince d’Arenberg, President of the Canal Company, and Sir Horace 
Pinching, late Director-General of the Egyptian Public Health 
Department, having been obtained. Two mosquito brigades were 
formed, one for the European and one for the native quarters of the 
town, and the oiling of all stagnant water practised once every weel^ 
Cess-pools were re-built and cellars filled up, with the result that 
within three months the mosquitoes were reduced to a negligible 
quantity, and mosquito nets largely dispensed with. Now, after two 
years, mosquitoes have become so rare in the town that they can be 
ignored, and malaria, though never very prevalent, has completely 
disappeared. But dengue fever has disappeared also, no case having 
been treated in Port Said since July, 1906. During the early part of 
that year, before the mosquito work began, dengue fever made its 
appearance as usual. Thirteen cases were treated in the hospital 
alone during April and May, and then as the mosquitoes disappeared 
the disease stopped and has not recurred since. In September, 190C, 
a severe epidemic raged throughout Egypt, beginning at Assouan and 
running rife in Cairo and Alexandria. It appeared in all the other 
towns, but Port Said and Ismailia remained free from it, no case 
occurring in either place. During the autumn of 1907 it again passed 
through Cairo and other parts of Egypt, but again Ismailia and Port 
Said escaped. Formerly the wards of the hospital in this town were 
full of cases of ‘ fever ’ during the summer months, but now the beds 
are used for other cases, which no longer contract fever although the 
mosquito nets have been removed. The extinction of the mosquito 
is greatly simplified in Egyptian towns owing to the dry summers, and 
the results can be easily watched. Port Said has a population of 
56,000, and Ismailia 10,000. The cost of the mosquito work in the 
former is sixpence per head of population per year, while in the latter 



195 


town it is nearly eighteenpence per head, owing to the extensive 
irrigation works which have to be regularly dealt with. 

It would seem, then, that the extermination of the domestic 
mosquito means the prevention of dengue fever, which, although not 
a very fatal disease, is one which causes endless misery in warm 
climates, and is as well a great hindrance to trade. 


LITERATURE 

1. Graham. Journal of Tropical Medicine, February i, 1903. 

2. Ashbirn and Craig. Philippine Journal of Science. B. Medical 

Scierces, Yol. II, No. 2, May, 1907. Quoted from Journal of the Royal 
Army Medical Corps, September, 1007. 


O 




197 


THE LIFE HISTORY OF TRYPANOSOMA 

LEW I SI 


BY 

J. E. SALVIN-MOORE 

PROFESSOR OF EXPERIMENTAL ANDjPATHOLOGICAI. CYTOLOGY, UNIVERSITY OF 

LIVERPOOL 

ANTON BREINL 

DIHECTOR OF THE RUNCORN RESEARCH LABORATORIES, LIVERPOOL SCHOOL OF 

TROPICAL MEDICINE 
AND 

EDWARD HINDLE 

DEMONSTRATOR IN THE RUNCORN RESEARCH LABORATORIES, LIVERPOOL SCHOOL 
OF TROPICAL MEDICINE 

(.Received for publication 5 May , 1908) 

The Trypanosomes, so far as at present known, are universally 
parasitic organisms inhabiting the blood and the body fluids of a 
variety of animals. In certain cases, the presence of trypanosomes 
produces the most marked pathological results. In others, the 
parasites are apparently quite harmless. Moreover, the same 
trypanosomes which are pathogenetic with respect to one animal, are 
often non-pathogenetic in the case of others. There is a tendency 
at present to attempt to draw a distinction in a classificatory sense 
between the so-called pathogenetic, and non-pathogenetic forms of 
trypanosomes. But even from the facts just referred to, it would 
seem to be clear that any such method of grouping can have but 
little real significance, and is more likely to entirely mislead enquiry 
than to throw any fresh light upon the singular and, morphologically 
speaking, closely knit group of organisms which the trypanosomes 
undoubtedly constitute. 

There appear, as a matter of fact, to be two main 
groups of problems connected with the trypanosomes at the 
present time. The first is constituted by our ignorance of the 
complete features of the developmental cycle of even any well-known 
and characteristic representatives of the group. The second by the 
present impossibility of determining to what group of non-parasitic 



iq8 


protozoa or protophytes the trypanosomes belong. There seems, 
however, to be little doubt that much of our present ignorance and 
confusion concerning the morphology and the life cycle of the 
trypanosomes has been due largely to an accident of technique, due 
in fact to the circumstance that the presence of trypanosomes can very 
readily be demonstrated by drying and staining the blood in which 
they are contained. It has so happened that by this method in its 
various forms, not only is the presence of trypanosomes demonstrated, 
but the preparations produced in this way are often extremely sharp, 
and beautiful to look at. It has consequently been only after a 
prolonged investigation of the effects produced upon such organisms, 
and on other forms of cells, by the process of drying, and by a careful 
comparison of the results obtained by this and other methods of 
fixation, th^t it has begun to be realised that dried preparations of 
cells, except for the purposes of demonstrating the presence of 
parasites, are generally as misleading as they are beautiful. 

We have referred to this matter in our former papers, and we may 
say that all our further acquaintance with trypanosome morphology 
indicates clearly that the process of drying is entirely destructive of 
the finer cytological details, and consequently that it is altogether 
inapplicable to investigations wherein a true conception of the normal 
features of trypanosomes, or indeed of any cells, is necessary. In 
consequence of these considerations, we have entirely abandoned the 
use of dried preparations, and have relied here, as in our former 
work, upon modifications of the various methods of fixation in 
common use among Cytologists, together with such modifications of 
the various staining methods as have been found necessary during 
the course of the work. The features of the developmental cycle or 
life history of T. lewisi , although remarkable, in reality only assume 
their true proportion when considered in conjunction with the facts 
relating to the development of other trypanosomes that have now 
been studied. 

For the sake of convenience, and for purposes of reference in 
subsequent portions of the present work, we shall in the first place 
briefly recapitulate our observations upon the developmental cycle in 
the case of T. gambiense and T. equiperdum . Before doing so, 
however, it is desirable to indicate the objects which we originally 
had in view in selecting the three forms, T. gambiense , T. equiperdum 



199 


and T. lewisi. Our primary intention with regard to T. gambiense was 
to ascertain what morphological results could be obtained with this 
form through the application of ordinary cytological methods in 
place of the usual drying process. But the subsequent results of this 
investigation were to reveal the existence of a life cycle among the 
parasites in the blood, which is definitely related to the alternating 
phases of presence and absence of trypanosomes in the circulation of 
infected rats.* The appearance of a cyclical metamorphosis among 
the trypanosomes in the blood indicated that the general conception 
of a special phase of their life history being definitely related to 
transference to another host (as is the case according to Schaudin 
with T. noctuae) might not be correct. Since, however, it is known 
that T . gambiense can be transmitted by the bites of tsetse flies, it 
was obvious that apart from investigations upon the transmitting 
insects, no ultimate conclusions could be arrived at with regard to this 
matter in the case of T. gambiense . 

There existed, however, in the disease Dourine, a trypanosome 
which under normal and natural conditions is not transmitted by any 
fly, or biting animal, but simply through contact. It was clear, 
therefore, that in this instance we had a trypanosome ljfe-history 
which was not normally complicated by the passage of the parasite 
through any intermediate host. Whatever life cycle T. equiperdum 
may possess, this cycle must be completed, and can be studied in the 
body of a single host. The acquisition of a knowledge of the facts 
relating to the life history of T. equiperdum^ the parasite of Dourine, 
was therefore of the first importance as a means of affording 
comparative material during a consideration of the significance of 
the features of the life cycle of T. gambiense in the blood. 

These two series of investigations in the case of T. gambiense 
and T. equiperdum having been undertaken, and both being related 
to parasites which produce violent and fatal maladies, it seemed 
further desirable to extend the investigation to some particular 
trypanosome which under normal circumstances belonged to the 
so-called non-pathogenetic forms. For this reason we selected 
T. lewisi . There was, however, another important point to be 
considered. T . lewisi can be transmitted from infected to non- 

• Salvin-Moore and Breinl. Annals of Trop. Med. & Parasitology, Vol, I, No. 3, 1907. 
f , „ Lancet, May 4, 1907. 



200 


infected rats by means of the rat louse, and consequently we had in 
this form of trypanosome a convenient object for investigating the 
changes which might take place among the trypanosomes when in 
the body of the louse, which here forms an intermediate host. 

P.assing now to the features of the developmental cycle in the 
parasites T. gambiense and T. lewisi in the blood, we find in the case 
of T. gambiense* injected into rats that the disease is marked by 
alternating phases of presence and absence of parasites in the 
peripheral circulation. If numerous preparations be made of the 
blood at short intervals during the whole course of the infection, it 
is found that at the time the parasites are increasing in number in 
the blood, rapid multiplication is going forward by means of 
longitudinal fission. Such fission is accompanied by amitotic division 
of the nucleus and the intra-nuclear centrosome (nucleolus, 
karyosome), as well as by amitotic division of the extra-nuclear 
centrosome (blepharoplast), and lastly by the development of a new 
flagellum, and the final splitting of the original trypanosome into two 
separate flagellated cells, each containing a nucleus, an intra-nuclear 
centrosome, an extra-nuclear centrosome, and a flagellum. 

Apart from the form of multiplication to which we have referred, 
no other form of reproduction takes place during the increase in the 
number of the parasites in the blood, and when we reach a point at 
or near the maximum number of trypanosomes in the circulation, the 
parasites cease to divide. At such periods it is found that in large 
numbers of them a stainable band develops from the extra-nuclear 
centrosome. This band extends, and finally becomes connected 
with the nucleus. It then breaks up and disappears. Subsequent 
to the development of the band, whether the trypanosomes again 
divide longitudinally, as in the case of T. lewisi , has not been 
ascertained. As we pass to those parts of the infection where the 
number of the parasites in the blood is falling, it is found that further 
rapid changes are taking place among the trypanosomes. The nuclei 
become more compact, vesicles appear in relation to them, and the 
nuclei, together with the vesicles, become separated from the outer 
portion of the cell, and enclosed by a delicate layer of cytoplasm. 
The remainder of such cells now disintegrates, and the composite 
body consisting of the nucleus, the vesicle, and a covering of 


* Salvin-Moore and Breinl, he. n't. 



201 


cytoplasm, becomes set free in the blood. These 'latent bodies ,’ as 
we have termed them, are eventually carried out of the peripheral 
circulation, and are subsequently to be found in large numbers in the 
spleen, the bone-marrow, and other organs. The process just 
described goes on until there may be no parasites to be found at all 
in the peripheral blood, but the latent bodies do not disappear, and 
after a time some of them grow larger in size, develop a new extra- 
nuclear centrosome (apparently from the division of the intra¬ 
nuclear centrosome), become flagellated, and finally gradually 
transform themselves into trypanosomes again. When this process 
has been completed, a similar cycle is passed through in relation to 
each alternating period of presence, and absence of the parasites in 
the blood, which the infected animal may present. It should be 
noted, however, that the cycle does not necessarily go forward at the 
same rate in all the trypanosomes present in an infected rat. All 
through the alternating periods there may be found a few trypano¬ 
somes at almost every stage of the cycle. 

Turning now to the development of 1 \ equiperdutn * it is found 
that in horses the infection presents the same sort of alternation of 
presence and absence of parasites in the blood that occurs during the 
infection of rats with T. gambiense , but in relation to the study of 
T . equiperdum in horses a difficulty presents itself. In such 
infections the parasites are so few, that it is practically impossible to 
obtain a sufficient number of them at the various periods of the 
curve of the infection for any adequate study. 

The same is the case when rabbits are infected with Dourine. We 
have therefore utilised rats, wherein the parasites multiply very 
rapidly, the features of the disease being as follows: — 

After injection no parasites appear until about the third day. 
They then multiply with extreme rapidity, and kill the animal in 
about four days after their first appearance in the blood. In rats, 
therefore, there is during an infection of Dourine only one 
developmental period, which is completed at, or about, the time of 
death of the infected animal. From the time of their first appearance 
the parasites multiply by longitudinal division; the features of this 
process being the same as those occurring during the multiplication 

* Salvin-Moore & JBreinl. On the Life History of Trypanosoma equiperdutn . 
Proc. Hoy. Sue., 1908, Vol. 60. 




202 


of the trypanosomes with an infection of T. gambiense. After the 
multiplication has proceeded for some time, two normal changes 
occur. The first is constituted by the budding off of a mass from 
the nucleus, the mass eventually disappearing altogether. The 
second obviously corresponds to the formation of the stainable band 
at a similar period of an infection with T . gambiense. In the case of 
T. equiperdum , this process proceeds by the production of a large 
bud, originating from the extra-nuclear centrosome. The bud 
rapidly increases in size, becomes detached, and passes towards the 
nucleus, with which it finally becomes definitely associated. 
Afterwards the trypanosomes again pass through divisions, and 
subsequently enter upon another change. They become altered in 
shape. The extra-nuclear centrosome becomes related to a long neck 
of protoplasm. A vesicle appears in relation to the nucleus. The 
extra-nuclear centrosome, together with the protoplasmic neck and the 
flagellum, becomes detached, and a large round body remains wherein 
a new extra-nuclear centrosome is apparent. From this a fresh and 
exceedingly delicate flagellum grows out. The extra-nuclear centro¬ 
some divides, and a second flagellum is produced. These large round 
double flagellated forms obviously correspond to the latent bodies of 
T. gambiense , but owing to the fact that the disease invariably kills 
the rats at or about this period, we have as yet been unable to follow 
completely the transformation of the large latent bodies into trypano¬ 
somes once more. The latent bodies seem, however, in the first place 
to divide up and to produce smaller forms, which latter probably 
correspond to those occurring in the life cycle of T. lewisi, as we 
shall see. 

T. lewisi is frequently found in the blood of wild rats in all parts 
of the world, but is rarely found in the blood of tame white rats. It 
is usually non-pathogenetic, and is relatively a large parasite. The 
morphology of the organism, and the various forms which it assumes 
in the blood, have been studied by several authors independently 
during recent years.* When a rat has become infected with T . lewisi , 

* Laveran and Mesnil. Ann. de l’lnstitut Pasteur, Vol. XV, No. 9, p. 673. 

Rabinowitsch and Kempner. Zeitsch. fur Hygiene und Infectionskrankheiten, Vol. 
XXX, p. 251. 

Wasielewski and Senn. Zeitsch. fur Hygiene und Infectionskrankheiten, Vol. 
XXXIII, p. 24b. 

Ward, J. MacNeal. Life History of Trypanosoma lewisi and Trypanosoma brucei. 
Journ. Infect. Diseases, Vol. I, No. 4. 

Prowazek. Studien uber Saugethiertrvpanosomen. Arb. a. d. Kaiser), Gesundheitsamtc, 
Vol. XXII, No. 2. 



203 


the parasites may be found in the blood in various forms at all periods 
of infection. Their development does not appear to occur in 
successive phases related to alternating presence and absence of 
trypanosomes in the blood, as is the case with T. gambiense and 
T. equiperdum. The elucidation of the developmental relationship 
of the various forms which thus exist together in the blood would at 
first sight seem to present a certain amount of difficulty, but in 
reality this difficulty is not so great as it appears. Thus the various 
authors who have already considered the subject are fairly well 
agreed with regard to the relationship in a developmental sense of the 
various forms one with another. Medium-sized parasites, such as 
those represented in fig. I, certainly give rise by growth to the large 
pointed types represented in figs. 11-18, so also these latter 
unquestionably pass into the still larger round and regular multi- 
nucleated masses, such as those represented in fig. 23. Such masses 
may again in turn be found in all stages of breaking up into smaller 
bodies, and this process of dissociation certainly produces the 
characteristic rosettes, and other forms of temporary association 
commonly met with (figs. 26-30). Longitudinal division among 
the medium-sized forms such as those represented in fig. 1, appears 
to be a rare occurrence, but that it does take place is indicated by such 
types as those represented in figs. 2-5, wherein the nucleus or the 
extra-nuclear centrosome, or both, have become divided, and a 
second flagellum has arisen. We have, however, ourselves not 
encountered the late phases of division in such forms of T. lewisi , and 
have consequently only been able to figure the early stages of the 
process. The forms such as those represented in fig. 1 appear 
to certainly arise from, and to merge into, forms such as those 
represented in figs. 37-38, and these undoubtedly in turn have arisen 
from the products of the dissociation of the large multi-nucleated 
masses. Thus we appear to have a cycle of development in the blood, 
which, starting from any particular type such as those represented in 
fig. 1, passes through the phases represented in figs. 12-19 m ^° 
the large forms represented in figs. 24-25. From this stage the cycle 
continues through stages such as those represented in figs. 30-38, 
and finally through growth and division the individual derivatives of 
the multi-nucleated masses pass back to the formation of types such as 
those originally chosen, as the starting point, and represented in fig. I. 



204 


That such a progressive development in the case of each individual 
really represents the course of the cycle in the blood, receives complete 
confirmation from the study of the various morphological changes 
which take place at the successive periods of the cycle; for these 
changes, as we shall see, correspond closely to the analogous changes 
which occur during the development of T. equiperdum and 
T. gambiense , that is to say, in forms where the successive stages are 
passed through approximately simultaneously by the majority of the 
parasites during the course of infection. 

The study of the morphology of T . lewisi may perhaps most 
simply be illustrated by taking in the first place examples such as 
those represented in fig. i. In this condition the cell is long and 
pointed at both ends. The extra-nuclear centrosome, which is large, 
lies at a considerable distance from the pointed extremity of the cell. 
The extra-nuclear centrosome stains very deeply with many forms of 
coloration, and can be seen during life as a highly refractive body. 
In various stained preparations the extra-nuclear centrosome appears 
to be always related to a vacuole, or space in the surrounding 
cytoplasm, and the flagellum may present various appearances in 
relation both to the vacuole and the extra-nuclear centrosome. The 
flagellum, which is a long stainable band, extends in a curved course 
over the whole length of the body, and projects at the opposite end 
as a whip-lash. It is enclosed in a thin expansion of the cytoplasm, 
forming the so-called undulating membrane. The flagellum generally 
ends in a small body, or bead, near the extra-nuclear centrosome (fig.i), 
but this is not always the case, for at times it certainly appears to run 
directly on to the extra-nuclear centrosome. When the flagellum is 
detached from the latter body, there can frequently be seen passing 
from the bead or thickening at the end of the flagellum, fine unstained 
strands which connect the bead with the extra-nuclear centrosome. 
The bead upon the end of the flagellum corresponds closely in 
appearance to the similar beads which are often found at the ends of 
the flagella among metazoan gametes, and such beads are in like 
manner often connected with the centrosomes by fine slightly staining 
strands. It would thus appear that so far as these structures among 
the trypanosomes can be directly homologised, the flagellum and its 
end-bead, together with the extra-nuclear centrosome, would 
correspond to the flagellum, bead, and centrosomes of many forms of 



205 


metazoan gametes. For this reason and others, to which we shall 
refer subsequently, we regard it as extremely misleading to name the 
end-bead a blepharoplast, and the extra-nuclear centrosome, a kineto- 
nucleus, for the end-bead (blepharoplast) does not present the 
relationships of a centrosome, or blepharoplast; while the extra- 
nuclear centrosome (kineto-nucleus) does so. Moreover, the extra- 
nuclear centrosome does not, so far as we are aware, present anything 
in common with a nucleus, except its capacity to. divide, and in this 
connection such a capacity amounts to nothing, for the capacity to 
divide is one which is, of course, shared by every known centrosome. 
The extra-nuclear centrosome is generally in the form of a thick rod, 
often slightly curved, and sometimes presenting the appearance of 
being divided in the middle. The division of the blepharoplast does 
not appear, however, to take place through any transverse separation, 
which such appearances might suggest 

The nucleus in T. lewisi lies relatively very near the end of the 
body from which the flagellum projects. In consists of an outer less 
stainable area, and a large inner much more darkly staining globe, the 
intra-nuclear centrosome (karyosome nucleolus). The outer portion 
of the nucleus is often very distinctly bounded, and in such phases of 
the development as those represented in figs. 14-19 might certainly 
be said to possess a membrane. During the phase of the development 
we are now considering, the cells do not present any very definite 
granules in the cytoplasm, which is seen, both under examination 
during life with a dark ground illumination, and after proper fixation, 
to consist of a fine protoplasmic foam bounded on the outside by a 
denser and homogeneous layer. 

Having thus briefly described the features of T. lewisi when in 
such a stage as that represented in fig. I, it will be most 
convenient in proceeding to describe the passage of such forms 
through the phases of the cycle we have already outlined, and to 
consider the various divisional and other phenomena as they occur in 
relation to this cycle. The form of trypanosome represented in 
fig. 1 passes by simple growth into the large forms represented in 
figs. 11-18, and all the intermediate stages can be readily found 
stretching from the morphological condition represented in fig. 1 
to that represented in fig. 17. Among such trypanosomes as those 
represented in figs. 8-17, two stages of metamorphosis are found to 



206 


occur. The first consists in the unequal budding of the intra-nuclear 
centrosome, so as to form what appears as a small nucleus, which 
becomes pushed off towards the free portion of the flagellum. This 
little mass, which consists of a small portion of the intra-nuclear 
centrosome and a small portion of the outer nuclear substance, appears 
subsequently to simply disappear. The process we have just 
described undoubtedly corresponds to the similar production of a 
degenerating nuclear bud in T. equiperdum . The second 
metamorphosis is constituted by the production of a body originating 
from the extra-nuclear centrosome. A portion of the substance of 
the extra-nuclear centrosome appears to pass round the adjacent 
vacuole, and to collect into a small mass on the side of the vacuole 
which faces the nucleus. This soon becomes completely detached, 
and passes away toward the nucleus through the cytoplasm. During 
its development the detached body becomes larger, and the outer 
portion of it stains less densely, but it is often possible to see a darkly 
staining bead at the centre of the growing mass (figs. 8-io). The body 
thus detached from the extra-nuclear centrosome may be found in all 
stages of transit from its original position to a close approximation to 
the nucleus (figs. 14-1O). Having reached this latter position, it appears 
to remain for some time unchanged. The process here described in 
T. lewisi obviously corresponds to the similar detachment of a portion 
of the extra-nuclear centrosome, and its subsequent passage to the 
nucleus, which we have described in T. equiperdum . It also 
undoubtedly corresponds to the formation of the stainable band 
stretching between the extra-nuclear centrosome and the nucleus 
during the life cycle of T. gambiense. The nucleus itself usually at 
this period begins to show signs of division. Such division which is 
represented in figs. 18-19 takes place in a typical amitotic fashion ; the 
intra-nuclear centrosome dividing like a drop as in T. gambiense and 
T. equiperdum , and the outer nuclear substance collecting round the 
two derivatives in the same manner. 

As in T. gambiense , T. equiperdum t and T. equinum i so also in 
T. lewisi we have been absolutely unable to observe anything during 
the division of the nuclei, or during any other periods, which in the 
remotest degree suggests the presence of chromosomes. During 
division of T. lewisi , the intra-nuclear centrosome first elongates, then 
becomes dumb-bell shaped, and finally assumes the form of two large 



globes widely separated from one another, and connected by a 
generally curved and tapering mass of substance which seems to have 
been simply drawn out between them. At the same time the outer 
nuclear substance collects about the diverging daughter elements, and 
finally separates along with them into two smaller masses, which, 
together with the new intra-nuclear centrosomes, eventually 
reproduce two complete and round nuclei, exactly like the parent 
nucleus only smaller. The process of nuclear division just described 
may be rapidly repeated; and at the same time the original 
trypanosome loses its characteristic form, and become rounded up so 
as to produce the well-known multi-nucleated masses such as those 
represented in figs. 12-25. I n some cases, however, when the nucleus, 
in a specimen such as that represented in fig. 19 has divided, the 
trypanosomes may become longitudinally split as in fig. 39, and in 
these cases the nuclei may at the same time travel towards the extra- 
nuclear centrosome so as to occupy the position represented in fig 39. 
The ends of these division products may become detached from one 
another, and a very curious appearance result, represented in fig. 39. 
We think that the features of this form of division at the period we 
are discussing deserves particular attention, for the appearances 
produced when it occurs are indistinguishable from the figures given 
by Prowazek, and interpreted by him as conjugation. 

We have, however, found nothing in relation to the nuclei, or any 
other structure in such cells, when in this condition, to suggest that 
the forms in question can be interpreted as conjugation. When such 
forms are produced, their future history appears to be this: Either 
the nuclei divide further, and the separation remains incomplete, the 
final product being one of the irregular multi-nucleated forms, or the 
fission is completed and the daughter cells, each after further nuclear 
divisions, produce fresh multi-nucleated masses. 

We have referred to this process because of its obvious bearing 
upon the interpretation to be put upon the identical figures given by 
Prowazek. Our observations indicate that it is relatively a rare 
method of procedure, the more normal processes being the 
multiplication of the nuclei and the rounding up of the trypano¬ 
some to produce eventually the multi-nucleated masses. During 
this period, i.e., the time and after the body becomes detached 
from the extra-nuclear centrosome the history of this latter 



208 


structure in T. lewisi is difficult to follow. It certainly often 
enters into close contact with the nucleus before division as in 
figs. 15,16, but it is frequently discernible after the nuclei have divided, 
as in fig. 20. In some of the resulting forms, moreover, when two, 
three, or four nuclei have been produced, the extra-nuclear-centrosomic 
derivative may sometimes still be observed lying between the nuclei, 
and apparently in close association with them. Whether the 
substance of the extra-nuclear-centrosomic derivative is directly 
absorbed by the nuclei, or merely disappears in the cytoplasm, the 
body in question sooner or later vanishes, and cannot be observed any 
further. The division of the nuclei of T. lewisi in the form we have 
just described is accompanied by the fission of the extra-nuclear 
centrosome, the fission of this latter body being generally 
accompanied by a movement towards the nucleus. It sometimes 
happens, however, that not only does the extra-nuclear centrosome 
move towards the nucleus, but the nucleus itself also moves towards 
the extra-nuclear centrosome. The advent of division of the extra- 
nuclear centrosome is marked by the development of the rod-like 
form into a flat disc, which perhaps through its thinness stains less 
darkly than the extra-nuclear centrosome when in a condition of rest 
(figs. 40-42.) The next phase is constituted by the collection of the 
staining material on opposite sides of the disc, and finally by the 
production in this way of two curved rod-like bodies on each side of 
the disc (figs. 41, 42). These new rod-like bodies constitute the new 
extra-nuclear centrosomes. They now rapidly diverge; it may be 
widely, showing at first a faint connection, which appears to be the 
remaining substance of the disc that has been simply drawn out This 
connection rapidly disappears, the resulting extra-nuclear centrosomes 
having then the same appearance as those in the parent form; but 
they are naturally smaller. During the nuclear division at this period 
which result in the production of the multi-nucleated masses, the 
division of the extra-nuclear centroseme does not, so far as we have 
been able to see, result directly in a division of the flagellum or the 
body attached to its proximal end. During such phases in T. lewisi, 
the original flagellum and its bead remain unaffected, and apparently 
do nothing. When the extra-nuclear centrosome has divided, as in 
fig. 44, it is often seen that a small body is closely attached to it, 
appearing as if it had been separated from the extra-nuclear 



2og 


centrosome. These little granules lie in the position from which new 
flagella finally arise, and it is consequently suggested that in T. lewisi 
the flagellum originates from a small fragment of the extra-nuclear 
centrosome, which becomes detached after the extra-nuclear 
centrosome has divided. This view of the method of procedure is 
further enforced by the fact that after the dissociation of the 
flagellum from the extra-nuclear centrosomes (which take place during 
the division of the latter bodies, see figs. 41-46), the original flagellum 
and its bead appears to be left, and is certainly finally shed in a 
degenerative condition, in the same way as the flagellum is cast off 
during the formation of the latent bodies in T. gambiense and 
T. equiferdum. From the vicinity of each of the new extra-nuclear 
centrosomes, and apparently from the granules budded off from these 
bodies, new and delicate flagella arise, and the multi-nucleated mass 
may assume in consequence appearances such as those represented 
in figs. 41-46. 

It will be seen that the features of the phase we have now 
described, that is, the production of the large pointed forms, the 
passage of an extra-nuclear-centrosomic derivative to the nuclei, the 
subsequent division of the nuclei, the formation of new extra-nuclear 
centrosomes, the degeneration and disappearance of the old 
flagellum and the formation of new flagella in association with new 
extra-nuclear centrosomes, certainly correspond in a biological sense 
with the phases we have considered and described in relation to the 
production of the latent bodies in T . gambiense and T. equiferdum. 
It would seem, indeed, that the multiplication of the nuclei in the large 
multi-nucleated masses of T. lewisi correspond to the division in 
T. equiferdum after the passage of the extra-nuclear-centrosomic 
derivative to the nucleus. The subsequent history of the multi- 
nucleated forms is equally interesting in this comparative aspect. The 
nuclei and the extra-nuclear centrosomes may become multi-nucleated 
till there are 10, 15, or more of each in a single mass. The flagella 
become distributed on the periphery of such masses, and the mass 
finally separates by forming either a mulberry-like aggregate of round 
flagellated forms, or the fission proceeds in a slightly different manner, 
and a curious group of somewhat elongated forms may be produced, 
as fig- 2Q. In all these resulting forms, whether elongated or round, 
the morphological conditions are quite different from those of the 



210 


characteristic trypanosome form. The nuclei occupy a more or less 
central position. The body of the cell is short (fig. 28), or actually 
round (fig. 31), and the long delicate flagellum is quite free. When 
such forms are elongated, as in fig. 27, the flagellum and the extra- 
nuclear centrosomes lie together on one side of the nucleus, and the 
flagellum passes away from the vicinity of the extra-nuclear 
centrosome in an opposite direction to that of the nucleus. These forms 
arising from the ultimate breaking up of the multi-nucleated masses 
are thus seen to possess all the morphological characteristics of the 
latent bodies, which are produced after the corresponding cycle of 
internal changes in the case of T. gantbiense and T. equiperdum. The 
derivatives of the multi-nucleated masses in T. lewisi thus correspond 
to the latent bodies of T. gantbiense and T. equiperdum . In T. lewisi , 
however, the subsequent history of the latent bodies is far more easy 
to follow than in any case which we have hitherto been acquainted. 
T. lewisi in this respect constituting an admirable example for the 
further study of this important phase, which is less easy to follow in 
the development of the pathogenetic forms to which we have referred. 

The changes which succeed in the small flagellated forms or latent 
bodies of T. lewisi are essentially similar to the transformation of the 
latent bodies of T. gambiense into the ordinary trypanosome form. 
The latent body elongates, the flagellum at first passing directly away 
from the surface of the body and in a direction opposite to that in 
which the nucleus lies in respect to the extra-nuclear centrosome 
(fig. 31). After a time the extra-nuclear centrosome migrates to one 
end of the cell body (fig. 32), and the flagellum is apparently drawn 
over the surface of the body after it. This portion of the flagellum 
which remains attached to the cell forms, as it were, the ‘ Anlagen * 
of the future undulating membrane. The further development is 
simple, the body elongating and enlarging into the ordinary 
trypanosome shape, as in figs. 33-38. When the form of small try¬ 
panosome, such as that represented in fig. 38, has been assumed, 
the cells again enter into division, as may be seen in figs. 34-37, and 
through the process of growth and further fissions gradually pass 
back again to the forms with which this description started (fig. 1). 

In briefly considering the foregoing observations upon the life 
history of T. lewisi , the most striking biological feature which emerges 
is the obvious similarity that exists between the successive phases 



211 


presented by T. lewisi , and the homologous phases occurring in the 
life cycles of T. gambiense and T. equiperdum . In each of these 
three cases the * trypanosome form ’ multiplies through fission until an 
interaction takes place between the extra-nuclear centrosome and the 
nucleus. This interaction may be succeeded again by simple fission, 
as in T. equiperdum. Possibly this is also the case in T. gambiense , 
while in T. lewisi it is followed by a series of rapid nuclear divisions 
resulting in the formation of the characteristic multi-nucleated masses. 
These differences, however, appear to be mere specific differences, of 
quite minor importance, which simply help to characterise in a specific 
sense the particular parasites we have considered. The divisions 
following the interaction between the extra-nuclear centrosome and 
the nucleus are, however, succeeded by a complete change of form, 
and by the assumption of the peculiar morphology of the round 
flagellated ‘latent body/ The fact that in T. equiperdum the latent 
bodies possess two flagella has probably a profound morphological 
significance, but it seems to be inappropriate at the present time, and 
in view of future work, to enter into a consideration of its actual 
significance. The details of the structure of the latent bodies, and 
their passage into the trypanosome form, is a matter which, although 
apparently simple in T. lewisi , is one which must receive further study. 
It is, for example, not at present clear in what way the two flagella of 
the latent body of T. equiperdum behave during this process. It 
becomes clear, when we consider the observations on the three forms 
to which we have referred, that during the life cycle in the blood the 
different phases in this cycle may become prolonged or shortened 
relatively with respect to one another. 

Thus the stage in T. gambiense, where one or two fissions possibly 
follow, the interaction between the extra-nuclear centrosome and the 
nucleus is prolonged in the case of T. equiperdum into a period where 
certainly several divisions take place, and this same period is again 
prolonged * and rendered specifically characteristic in the case of 
T. lewisi by the production of the large multi-nucleated masses. 

In this connection it seems also to be a very striking fact that, 
whereas in the pathogenetic forms T. gambiense and T. equiperdum 
the phases of the life cycle as they appear among the trypanosomes 
do so nearly simultaneously among all the parasites existing 
in the blood at a particular time, and thus mark successively 
p 



212 


the stages of the infection, in the non-pathogenetic form 
T. lewisi all the stages of the life cycle may be present and 
represented by different parasites which are found in the blood at the 
same time. This difference is perhaps what might have been 
expected Such forms as T. lewisi are usually present in the animals 
they inhabit in large numbers for weeks, or even months; whereas 
among the pathogenetic varieties the parasites are numerous in the 
blood for only relatively short periods, the phases of the life cycle 
being here apparently adapted to the varying conditions of the host. 

In this way we find that the parasites in such forms either 
multiply without limit, and by their action rapidly kill the host, or they 
periodically disappear from the altered blood in the form of latent 
bodies, and only reappear, it may be, after a very considerable time. 

We have referred, in dealing with T. gambiense and 
T. equiperdum , to the fact that we have been quite unable to make 
anything of the arbitrary distinction which has come into vogue since 
the publications of Schaudinn between the so-called males, females, 
and any different forms. These seem to us to be either mere 
varieties of size, or, where morphological distinction is obtained, 
examples that have been taken from different parts of the life cycle. 

The same results in relation to this matter have been enforced by 
the study of T. lewisi. Moreover, the terms male and female have, 
biologically speaking, always a strict and obvious reference to two 
varieties of cells which conjugate, or gametes, and to use terms of this 
type in reference to mere varieties of size, or to the morphological 
characters of different phases in a life cycle where no ordinary 
con jugation has hitherto been found, seems to us to be in the highest 
degree misleading and erroneous. 




214 


PLATE II 
T. Icwisi 

Figs. 1-6. Stages of rest and division of the nucleus, the extra- 
nuclear centrosome, and the development of new flagella. 

Fig. 7.—Stage in the division of the nucleus. 

Fig. 8.—Formation of a small nuclear body which is thrown off from 
the nucleus and production of a large mass from the 
extra-nuclear centrosome. 

Figs. 9, 10, II.—Further stages in the development of the body 
derived from the extra-nuclear centrosome, and its passage 
towards the nucleus. 







2l6 


PLATE III 
T. lewisi 

Figs. 12, 13, 14, 15, lO, 17.—Stages in the passage of the body 
developed from the extra-nuclear centrosome towards the 
nucleus. Figs 12 and 17 show the degeneration of the 
small mass detached from the nucleus. 

Figs. 18, 19.—Stages of the division of the nucleus. 

Fig. 20.—Partly rounded form with two extra-nuclear centrosomes, 
and the body derived from the extra-nuclear centrosome. 

Fig. 21.—Rounded mass produced after the division of the nucleus, 
and extra-nuclear centrosome. 

Fig. 22—Division of a nucleus in a large rounded form. 




rfxx/re.BrwiL ^ 




PUt.ui 








2l8 


PLATE IV 
T. lewis i 

Fig. 23.—Large form showing two nuclei and two extra-nuclear 
centrosomes. 

Figs. 24, 25.— Large multi-nucleated masses, fig. 24 showing division 
of one of the nuclei. 

Figs. 26, 27, 28, 29, 30.—Breaking up of the large multi-nucleated 
masses into forms equal to the latent bodies of T. 
gambiense and T. equiperdum . 

Figs. 31, 32, 33, 34, 35, 36, 37, 38.—Stages of transformation of the 
latent bodies into ordinary trypanosomes. 



c Woot-e.BreijJ Hrndla. 


P late IV. 



V d ’?! 







220 


PLATE V 
T. lewisi 

Fig. 39 -—Division which at first sight suggests an act of conjugation. 

Figs. 40, 41, 42, 43.—Forms showing details of the division of the 
extra-nuclear centrosome. 

Fig. 44.—Form showing detachment of small bodies from the extra- 
nuclear centrosome. 

Figs. 45, 46.—Details of the origin of the flagella. 

Fig. 47.—Division of the nuclei in a multi-nucleated mass. 

Figs. 48, 49.—Unusual division of the nuclei. 






221 


NOTES ON THE EFFECTS OF THERA¬ 
PEUTIC AGENTS ON TRYPANOSOMES 
IN RESPECT TO (a) ACQUIRED RESIS¬ 
TANCE OF THE PARASITES TO THE 
DRUG, AND (b) CHANGES IN VIRULENCE 
OF THE STRAINS AFTER ESCAPE FROM 

THE DRUG 


BY 

BENJAMIN MOORE, M.A, D.Sc. (R.U.I.) 

JOHNSTON PROFESSOR OF BIO-CHEMISTRY 

MAXIMILIAN NIERENSTEIN, Ph.D. (Berne) 

INTERNATIONAL RESEARCH FELLOW 

AND 

JOHN LANCELOT TODD, B.A., M.D. (McGill), M.R.C.S. 

PROFESSOR OF PARASITIC PROTOZOOLOGY, McGILL UNIVERSITY, MONTREAL 

(From the Departments of Bio-Chemistry and of Tropical Medicine , 
University of Liverpool) 

(Received for publication u May, 1908) 

A. DEVELOPMENT OF DRUG-PROOF STRAINS 

It is a well-established fact in the therapeutics of trypanosomiasis 
that, with each fresh recurrence of the parasites in any case, these 
become more resistant to the drug employed, and more difficult to 
drive out of the peripheral circulation. 

This is not only the fact in any given case, but also as was first 
shown by Ehrlich* in the case of rats treated by atoxyl; the parasite 
itself, apart from the individual host, becomes resistant to the drug, 
and, when passed to fresh animals of the same species, reaches finally 
a condition in which it is resistant or 1 fast 1 to the particular drug. 
This condition Ehrlich expressed by the term ‘ Atoxyl-fest ’ in the 
case of trypanosomes which had become refractory to the action of 
atoxyl. 


Berl. Klin. Wochensch., 1907, p. 33. 



222 


We have observed similar effects, and they have also been noted 
by Plimmer and Thomson* in the case of atoxyl. Ehrlicht also found 
a developed resistance in the case of certain of the anilin colours used 
by him as trypanocides, the colours experimented with being para- 
fuchsin, trypan-red and trypan-blue. The special point which we 
believe is shown in regard to this resistance by the experiments here 
recorded is that it only holds for the species of animal in which it has 
been established, and disappears when the fast strain of trypanosomes 
is passed into another species of animal. 

T. brucei was used in the experiments, and two distinct strains 
were used: ( a ) a strain which was atoxyl-fast in mice, for which we 
are indebted to the kindness of Prof. Ehrlich, and ( b ) an atoxyl-proof 
strain in the rat, obtained by ourselves after eight relapses. The 
results with the first of these strains are given in Table I, those with 
the second strain in Table II. 


Table I 


Strain atoxyl-fast in mouse as shown by first three experiments, 
but not fast in rat and dog as shown by subsequent experiments. 


Experiment 
i. Mouse (84) 


2. Mouse (85) 


Treatment 


Duration of 

life Examinations 


0*5 c.c. of 2 per cent. 8 days 
acetylated atoxyl] 
repeated three times 


As above 


7'5 da >' s 


Trypanosomes seen 
all the time, appa¬ 
rently diminished 
after second injec¬ 
tion 

As above 


3. Mouse (92) 

4. Rat (265) 


0-5 c.c. of 5 per cent, 
solution of atoxyl 


6 days 
19 days 


Trypanosomes dis¬ 
appeared and re¬ 
appeared in twenty 
days 


5. Rat (266) 

6. Dog (312) 


As above 


24 days 


10 c.c. of 10 per cent. 29 days 
solution of acety¬ 
lated atoxyl in two 
injections 


Trypanosomes dis¬ 
appeared and re¬ 
appeared on twenty- 
sixth day 


* Proc. Roy. Soc.,'Vol. 79B, 1907, p. 504. 
t Loc. cit. 

J A supply of tliis substance was kindly sent us by Professor Ehrlich. 



223 


Table II 


Strain atoxyl-fast in the rat as shown by first three experiments, 
but amenable to atoxyl in the mouse and dog as shown by the other 
experiments. 


Experiment 


Treatment 


Duration of 

life Examinations 


i. Rat (289) 


1 c.c. of 5 per cent, 
atoxyl 


9 days Trypanosomes were 
found on all but 
one day 


2. Rat (292) As above 


11 days Trypanosomes present 
all the time 


3. Rat (298) 


1 c.c. of 5 per cent. 10 days 
atoxyl, twice re¬ 
peated 


Trypanosomes present 
all the time 


4. Mouse (94) 


0 5 c.c. of 2 per cent. 23 days 
acetylated atoxyl 
given three times 


Trypanosomes dis¬ 
appeared in twenty 
days 


5. Mouse (97) As above 


26 days Trypanosomes dis¬ 
appeared in twenty- 
three days 


6. Dog (316) 


10 c.c. of 10 per cent. 19 days 
acetylated atoxyl 


Trypanosomes dis¬ 
appeared and re¬ 
appeared in fourteen 
days 


B. VARIATIONS IN VIRULENCE IN FRESH ANIMALS OF STRAINS 
WHICH HAVE REAPPEARED AFTER TREATMENT WITH 
DIFFERENT DRUGS 

In our work on the treatment of Ngana ( T . brucei) by atoxyl and 
mercury salts in rats, we have observed that when the parasites had 
reappeared after atoxyl treatment, and were then passed on into a 
fresh animal, they had acquired increased virulence in the process and 
caused death more rapidly than when passed on from animal to 
animal without any treatment. Accordingly a series of experiments 



224 


with animals infected with T. brucei , T. gambiense and T. dimorphon 
was instituted specially to make observations on this point. In the 
case of the infections with T. brucei it was found that the virulence 
increased after attack with atoxyl, but was diminished after trypan- 
red. A slight increase in virulence was seen in the few experiments 
done with T. dimorphon , and a slight decrease in the case of 
T. gambiense , but the number of experiments and alteration in the 
virulence are both too small to warrant a definite conclusion in the 
case of these two species. The number of experiments with T. brucei 
is larger, and we are of opinion that this parasite after escape from 
atoxyl is, in the majority at least of the cases, more virulent than 
before treatment. 

Previous experiments, without treatment with any drug, gave as 
the average duration of life after appearance of infection in the case 
of each strain as follows : — 

T . brucei 6 3 days Average of 146 rats 

T. dimorphon 17*6 „ ,, 56 „ 

T . gambiense 287 „ ,, 36 „ 

No sudden alteration in virulence was found in the case of any of 
the strains. This had been specially watched for in the case of the 
strain of T. brucei , which had been under observation from 
4th November, 1906, till 16th July, 1907; as stated, no increase of 
virulence was seen in the strain passed in the usual way from animal 
to animal without coming under the influence of any drug. 

In order to infect the animals, an injection subcutaneously of 
0 4 c.c. of infected blood, containing 6 to 8 parasites to the field 
(Zeiss, ocular 4, objective DD), was given in each case. 

The results are shown in the following tables, which show the day 
of reappearance of trypanosomes in the peripheral circulation after 
their disappearance under the influence of the drug mentioned in each 
case, the number of animals then infected by the recurrent strain from 
the drug, the duration of life under this strain, and the increase in 
virulence marked -1-, or decrease marked — . 



22 5 


Table III 

Virulence of T. brucei recurrent after atoxyl treatment. 

No. of animals infected Average duration Change of 


Day of relapse 

with recurrent strain 

of life in davs 

virulence 



(Untreated rats 146) 

( 6 * 3 ) 


23rd 

day 

4 rats 

4-6 

+ 17 

29th 


7 - 

5 *^ 

+ i ‘3 

19 th 


2 „ 

6*o 

+ 0-3 

24th 


4 .. 

3-8 

+ 2-5 

26th 


3 .. 

6*8 

- 0-5 

38th 


7 .. 

4*2 

+ 2-1 

25th 


2 „ 

5 *o 

+ 1-3 

19th 


3 

6*o 

+ 0-3 

29th 


5 » 

7*5 

— 1*2 

21S 


8 „ 

4’5 

+ i-8 

35th 


6 „ 

4 *o 

+ 2-3 

31st 


3 .. 

6*8 

- o -5 


Taking the averages of the whole numbers of untreated and 
treated rats, we have : — 

Untreated rats 146 Average duration of life 6*3 days 

Post-atoxyl-treatment rats 54 ,, ,, ,, 5*1 „ 

Mean increase of virulence 1*2 „ 

The same strain of T. brucei treated with trypan-red in three 
experiments in which 11 rats were infected from recurrences after 
this drug, showed on the other hand a considerable decrease in 
virulence of the strain, as shown in the following table. 


Table IV 

Virulence of T. brucei recurrent after trypan-red treatment. 


Day of relapse 

No. of animals infected 
with recurrent strain 

Average duration 
of life in davs 

Change of 
virulence 

7th day 

(146 rats untreated) 
4 rats 

( 6 - 3 ) 

15 

- 97 

nth „ 

3 - 

9 

- 2 7 

5 t h .. 

4 - 

11 

- 47 


Untreated rats 146 Average duration of life 6*3 days 

Post-trypan-red-treatment rats 11 ,, „ „ n g ,, 

Mean decrease of virulence 5*6 ,, 







226 


The strain of T. dimorphon , after treatment with atoxyl, gave 
the following results in five experiments with 18 rats:— 

Table V. 

Virulence of T. dimorphon recurrent after atoxyl treatment. 

No. of animals infected Average duration Change of 
Day of relapse with recurrent strain of life in days virulence 

(Untreated rats 56) (17*6) 

29th day 5 rats 14*5 + 3*1 

42nd ,, 2 „ 190 — 1*4 

34th „ 4 „ II-O + 6-6 

28th „ 3 ,, 19*0 — 1*4 

26th ,, 4 „ 16-0 -f i-6 

Untreated rats 56 Average duration of life 17*6 days 

Post-atoxyl-treatment rats 18 „ „ „ 15*3 „ 

Mean increase in virulence 2*3 „ 

In four experiments with the strain of T gambiense , in which 20 
rats were used, the following decrease in virulence was observed, but 
as stated above a much larger number of experiments would be 
required in order to definitely decide the point, for this much slower 
species of trypanosome: — 

Table VI 

Virulence of T. gambiense recurrent after atoxyl treatment. 


Day of relapse 

No. of animals infected 
with recurrent strain 

Average duration 
of life in days 

Change of 
virulence 

59th 

day 

(36 rats untreated) 

6 rats 

(287) 

32 

“ 33 

62nd 

n 

7 - 

30 

- i-3 

54th 

it 

4 » 

39 

- 103 

68th 

11 

3 - 

31 

2*3 


Untreated rats 36 Average duration of life 287 days 

Post-atoxyl-treatment rats 20 ,, „ ,, 32 5 ,, 

Mean decrease in virulence 3*8 ,, 

It may, perhaps, for clearness be emphasized that none of the 
animals mentioned in the Tables (III to VIII) received any treat¬ 
ment whatever. The comparisons are entirely of the virulence of 
the strains in what might be described as their natural condition, 
and when they have reappeared after a drug and are passed into a 
fresh host of the same animal where they are not further treated. 



227 


OBSERVATIONS ON THE ACIDITY 
AND ALKALINITY OF THE BLOOD 
IN TRYPANOSOME INFECTIONS 


BY 

M. NIERENSTEIN, Ph.D. 

J. W. GARRETT INTERNATIONAL FELLOW AND RESEARCH DEMONSTRATOR OF 
THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE 

(From the Runcorn Research Laboratories) 

(Received for publication 12 May , 1908 ) 

Yakimoff 1 in his publications dealing with the changes of blood 
during trypanosome infection states that the alkalinity of blood 
decreases as the disease progresses. He used for his estimations 
von Limbeck’s 2 method, whereby the serum alkalinity is measured 
against litmus. This method, however, does not indicate the true 
alkalinity of the serum, as the results are affected by the increase of 
the acidity and the carbon dioxide present in the blood 

A method which practically eliminates these errors has been 
described recently by Moore and Wilson, 3 who estimate the 
alkalinity of the ash after incineration of the blood. We were able to 
show, with their method, that during infections with T. brucei and 
T. equiperdum the acidity of the blood serum increases, whereas the 
alkalinity of the blood apparently remains constant. 

B. Moore’s and F. Wilson’s technique was adopted, phenolph- 
thaleine being used as an indicator for alkalinity, and dimethyl-amido- 
azo-benzol (referred to as dimethyl for brevity) for acidity. In 
addition titrations were made with Congo red, as this does not indicate 
organic acids such as amido acids. 4 

It is remarkable that in cases where both indicators were used, 
the acidity against Congo red was lower than against phenolphthaleine. 
This difference becomes more marked as the infection progresses, a 
fact which seems to suggest that trypanosome infection causes an 
increase of amido acids in the blood. 

To eliminate the error which might be produced by the CO a in 
the breath, to which Moore and Wilson allude in their paper, special 

Q 



228 


precautions were taken. The mouth-part of the pipette contained 
KOH between two layers of cotton wool, so that all the CO a of the 
breath was absorbed by the potassium hydroxide. 

The blood was collected in small test tubes and left standing over 
night in an ice chest; then the serum was separated from the clot and 
used for the estimation of acidity. For the estimation of the 
alkalinity, the blood was collected in a platinum crucible and 
incinerated. All the glass vessels used in this work were immersed in 
strong hydrochloric acid for three days, and then in distilled water for 
the same length of time, so as to avoid the error which might be 
produced by the alkalinity of the glass. 

Experiment No. 303.—Rabbit, male, inoculated on 10th February, 1908, with 
T. equiperdum. On 12th February the acidity was 0-03015* (phenolphthaleine) ; on 
24th February the acidity had increased to 0-03255 (phenolphthaleine), 0-03195 
(congo red), the alkalinity was 0-02895. The animal was frequently examined and 
kept under observation until 7th April, when the acidity had reached 0-03495 
(phenolphthaleine), 0*03345 (congo red), and the alkalinity 0*03015. 

Experiment No. 305.—Rabbit, female, inoculated on 10th February, 1908, with 
T. equiperdum. On 24th February the acidity was 0-03285 (phenolphthaleine), 
0-03225 (congo red), and the alkalinity 0*02865. On 7th April the acidity had 
reached 0-02865 (phenolphthaleine), 0-03405 (congo red), and the alkalinity 0-02925. 
It is interesting to note that on 31st March the acidities against congo red and 
phenolphthaleine were the same, 0*03465. 

Experiment No. 307.—Guinea-pig, female, inoculated on 24th February, 1908, 
with T. brucei. The first alkalinity and acidity estimations were made on 26th 
February, when the acidities were found to be 0*03105 (phenolphthaleine), 0*03075 
(congo red), and the alkalinity 0*03045. On nth March the animal was found to 
be swarming with parasites, and the acidity estimation on 14th March gave 
0*03435 (phenolphthaleine), 0-0327 (congo red), and the alkalinity 0*03000. The 
animal died on 18th March of typical trypanosomiasis. The acidity estimation 
made twenty minutes after death gave 0*03615 (phenolphthaleine), and 0*03480 
(congo red). 

Experiment No. 309.—Guinea-pig, female, inoculated on 24th February, 1908, 
with T. brucei. The first acidity estimation was made on 26th February, and found to 
be 0*03195 (phenolphthaleine), 0-03175 (congo red), and the alkalinity 0*03030. The 
animal showed trypanosomes on the same day. On 7th April the acidity had 
reached 0-03465 (phenolphthaleine), 0*03450 (congo red), and the alkalinity 0*02925 ; 
the animal was then heavily infected. 

Experiment No. 310.—Guinea-pig, female, inoculated on 24th February, 1908, 
with T. brucei. On 26th February the acidity was 0*03225 (phenolphthaleine), 
0 03210 (congo red), and the alkalinity 0-02895. On 8th March the acidity was 
0*03405 (phenolphthaleine), 0*03300 (congo red). A fall m the acidity was noticed on 
14th March. The acidity reached 0*03x95 (phenolphthaleine), and the same also for 
congo red. Afterwards the acidity started slowly to increase and became on March 
23rd 0*03510 (phenolphthaleine), 0*03375 (congo red), and the alkalinity 0*02985. 
On 7th April the acidity had reached 0-03585 (phenolphthaleine), 0*03495 (congo 
red), and the alkalinity 0*03045. 

* Expressed in fractions of Normal. 



229 


Experiment No. 31 i. —Guinea-pig, male, inoculated on 24th February, 1908, 
with T. brucei. On 24th February the acidity was found to be 0*03245 (phenolph- 
thaleine), 0-03225 (congo red), and the alkalinity 0*03000. The last estimation was 
made on 17th March, and the animal died on 19th March. The animal was then 
slightly infected, and the acidity found to be 0*03300 (phenolphthaleine), 0*03255 
(congo red), and the alkalinity 0*03000. The cause of the death of this animal was 
pneumonia. 

Experiment No. 314.—Rabbit, female, inoculated on 14th Maich, 1908, with 
T. brucei. On the day of inoculation the acidity was found to be 0*03075 
(phenolphthaleine), 0*03030 (copgo red), and the alkalinity 0-02850. On 23rd March 
the acidity was found to be 0*03300 (phenolphthaleine), 0*03195 (congo red), and the 
alkalinity 0-03060. On the same day, three young ones were born which soon died. 
The acidity on 26th March had dropped down to 0*03240 (phenolphthaleine), 
0*03225 (congo red), and the alkalinity 0*02850. From this day the acidity 
increased slowly, and on 7th April was 0*03345 (phenolphthaleine), 0-03285 (congo 
red), and the alkalinity 0*03015. The animal died on 9th February. This animal 
was positive all the time, starting from 17th March. 

Experiment No. 351.—Rabbit, female, inoculated on 14th March, 1908, with 
T . brucei. The acidity was then 0*03120 (phenolphthaleine), 0*03075 (congo red), and 
the alkalinity 0*02940. On April 7th the acidity had reached 0*03375 (phenolph¬ 
thaleine), 0*03315 (congo red), and the alkalinity 0*02910. 

Experiment No. 316.—Rabbit, male, inoculated on 14th March, iqo8, with 
T . brucei , showing an acidity of 0*03180 (phenolphthaleine), and 0*03075 (congo red), 
and an alkalinity of 0*02880. When examined on 31st March, the acidity had 
reached 0*03375 (phenolphthaleine) 0*03210 (congo red), and the alkalinity 0*03000. 

Experiment No. 317.—Guinea-pig, female, inoculated on 14th March, 1908, 
with T. brucei. The acidity was then 0*03150 (phenolphthaleine), 0*03105 (congo 
red), and the alkalinity 0*02895. The animal died on 8th April of typical 
trypanosomiasis On 7th April the acidity had reached 0*03585 (phenolphthaleine), 
and 0*03360 (congo red), when the alkalinity was 0*02850. 

I should like especially to draw attention to this experiment, in 
which both the increase of the total acidity and also that of the 
amido acids is very marked. 

The following table gives in full the changes in acidity and 
alkalinity during an experimental infection : — 


Experiment No. 304.—Rabbit, male, inoculated 10th February, 1908, with 


T . cquiferdum. 


Date 

12/2/08 

Acidity to 
phenolphthaleine 
0*03135 


Acidity to 
coneo red 

Alkalinity to 
dimethyl 

14/2/08 

0*03195 


— 

— 

18/2/08 

0*03225 


_ — 

0*02895 

20/2/08 

0*03285 


0*03195 

0*02880 

24/2/08 

0*03240 


0*03210 

0*02880 

27/2/08 

0-03240 


0*03225 

— 

2/3/08 

0*03255 


0*03210 

0*02895 

4/3/08 

0*03300 


0-03210 

002865 

8/3/08 

0*03255 


0*03225 

— 

10/3/08 

0-03285 


0*03210 

0*02895 

14/3/08 

003255 


0*03210 

0*02865 

1 7/3/08 

0-03255 


0*03255 

0*02880 

19/3/08 

0*03285 


0*03225 

0*02910 

*3/3/08 

0*03315 


0*03225 

0*02985 

1.—Trypanosomes examinations 

were 

made daily. 

The animal died on 

when the post-mortem showed a 

more 

or less marked trypanosomiasis. 







230 


The following table shows the acidity and alkalinity of normal 
rabbit serum:— 


Acidity to Acidity to Alkalinity to 


Sex 

F. 

phenolphthaleine 

0-03175 

Congo red 


dimethyl 

M. 

0*03225 

— 


— 

F. 

0-03075 

— 


— 

F. 

0-03180 

— 


— 

F. 

0*03075 

— 


-- 

F. 

0*03120 

— 


— 

M. 

0*03090 

— 


— 

F. 

0*03120 

— 


— 

F. 

0-03015 

— 


— 

F. 

0*03175 

— 


— 

F. 

0*03075 

— 


— 

F. 

0*03120 

— 


— 

M. 

0*03015 

— 


— 

F. 

0*03150 

— 


— 

F. 

°‘°3°7S 

0*03075 


0-02895 

M. 

0*03175 

°'°3 io 5 


0*02850 

F. 

0*03075 

0*03120 


0*02895 

F. 

0-03195 

0*03060 


0*02955 

F. 

0-03175 

0*03030 


0*02865 

F. 

0-03050 

0*03050 


0*03045 

F. 

0*03120 

0*03050 


0-03000 

M. 

0*03030 

003015 


0-02880 

F. 

0*03075 

0*03030 


0*02850 

M. 

0*03180 

0-03075 


0-02880 

F. 

003120 

0*03075 


0*02940 


Average 0-03115 

Average 0-03064 

Average 0*03025 


Conclusions. 

I. It is evident, that in experimental trypanosomiasis infection 
( T . brucei and T. equiperdum ), the acidity of the blood increases. 

II. The increase of the acidity is probably due to the production 
of amido acids through, or by the trypanosomes, i.e., the acids might 
be either secreted by the parasites or produced by the action of the 
parasites on the proteins of the blood serum. In the latter case, the 
amido acids would be broken up through hydrolysis from the proteins 
into simpler polipeptids. 

III. It is possible that the increase of acidity might be of 
assistance in the diagnosis of a typical case of trypanosomiasis, where 
the parasites have disappeared for some length of time from the 
blood circulation. 








231 

IV. These experiments suggest that in trypanosome treatment 
effort should be made to neutralise the increased acidity of the blood, 
as this might prove of additional assistance in making the blood a 
less favourable medium for their development. 


LITERATURE 

1. Jakimoff, W. L. K woprossu ob ismeneniach krowi srhiwotuich pri 

experimentalnich trypanosomach (Russian), St. Petersburg, 1907, pg. 18. 

2. Hammersten, O. Lehrbuch der physiologischen Cheinie (1907), p. 223, where 

the whole literature on the subject is to be found. 

3. Moore, B. and Wilson, F. A clinical method of Haemalkalimetry, etc. Bio- 

Chemical Journal, Vol. I, p. 297. 

4. Meyer, V. and Jacobson, P. Lehrbuch der organischen Chemie (1907). Vol. 

I, p. 702 . 







233 


CONTRIBUTIONS 

TO THE MORPHOLOGY AND LIFE 
HISTORY OF PIROPLASMA CANIS 


BY 

ANTON BREINL, 

DIRECTOR OF THE RUNCORN RESEARCH LABORATORIES, LIVERPOOL SCHOOL OF 

TROPICAL MEDICINE 

AND 

EDWARD HINDLE, 

DEMONSTRATOR AND RESEARCH ASSISTANT (<F THE KUNCoKN RESEARCH 

LABORATORIES 

(Received for publication 13 May, 1908; 

Piroplasma canis was discovered by Piana and Galli-Valerio in 
the year 1895, 20 and, on account of its wide distribution and the ease 
with which experimental infection can be transmitted from dog to dog, 
this parasite has been the subject of extensive study. It seems 
unnecessary to give a complete review of the literature, especially 
since Nuttall and Graham-Smith, 1719 and more recently Christophers, 3 
have given fairly complete bibliographies. 

Additional interest has been attached to Piroplasmata in general 
since the appearance of Schaudinn’s 21 work, in which he mentions 
(p. 428) Kossel's and Weber’s observation, and suggests that 
Piroplasma may pass through a life cycle similar to that of 
Halteridium. However, very little evidence in support of this 
hypothesis has been brought forward by later workers. 

The parasite, on which the following observations were based, was 
obtained from Professor Uhlenhuth, of Berlin, to whom we have 
pleasure in expressing our indebtedness. The strain has been kept 
going in pups and dogs by means of simple inoculation. In our hands 
it has shown itself very virulent, even in the case of full-grown dogs, 
and no animal survived the infection. Young dogs showed parasites, 
in scanty number, two to three days after an intra-peritoneal injection 
of 1 to 2 c.c. of heavily-infected blood; their number increased very 
slowly during the following 24 hours; after this a rapid increase in 
the number of parasites set in. Within 40 hours after the first 



234 


appearance of the parasites, the peripheral and especially the blood of 
the organs was usually teeming with them, and the animal succumbed 
to the infection. The course of the disease in full-grown dogs was 
somewhat modified, as the parasites only appeared after a prolonged 
incubation period, and never in such large numbers as in young 
animals. 

The clinical feature of this disease has been dealt with in full by 
previous workers. It is a noteworthy fact that in nearly all our cases 
haemoglobinuria was more or less pronounced, the urine being 
frequently of a dark port wine colour. Only in very few cases was 
jaundice well marked. 

Technique. All our observations were made on wet films using 
Breinl’s methods. The blood-smears were fixed in strong Flemming’s 
solution, and afterwards stained with safranine and methylene blue, 
according to Breinl’s method, or his modification of Heidenhain, 15 
using as counter-stain a dilute solution of Bordeaux red. By this 
means the cytological details of the parasites were well preserved, an 
attainment which is impossible by any dry film method. 


EARLY FORMS OF PARASITES IN THE BLOOD 

The early forms in the blood are usually very large and irregular, 
frequently exhibiting pseudopodia of varying form and size (figs. 1-5). 
Some of these processes are so fine that they simulate flagella, and at 
times small particles of protoplasm appear to become detached; but 
in most instances a very fine band may still be detected connecting 
these little masses with the parasite. The protoplasm consists of a 
fairly coarse spongioplasm (‘ Schaumplasma ’), containing fine, bluish 
staining granules, embedded in its substance. At this stage the 
parasites usually possess a single nucleus, in the form of a small, 
dense, darkly staining mass of chromatin, which is sometimes 
surrounded by a vacuole, filled with lightly staining substance. The 
division of these forms is by simple fission. The nucleus of the 
mother cell elongates, and afterwards separates into two halves which 
move further apart; meanwhile the parasite itself increases in size, 
and eventually divides into two daughter cells (figs. 2-4). This 
process goes on very rapidly. 

In the early forms very rarely a second smaller chromatic mass is 



2 35 


present (fig. 5), which may be connected with the main nucleus by a 
fine darkly staining line. If Breinl’s stain is used this smaller nucleus 
usually takes a dark purplish-blue colour, whereas the nucleus stains 
dark red. This difference is noticeable, however, only in well stained 
specimens. 

The division of the later forms proceeds in a different way. The 
small nucleus together with the chromatic line usually divides first. 
A median cleft afterwards appears, which extends between the two 
small nuclei. In the meanwhile the large nucleus elongates, and 
finally separates into two equal halves. The two daughter cells then 
become separate (figs. 6-9). This division results in the formation of 
two pear-shaped parasites. 

At this stage of the infection, round parasites now and again 
reproduce by budding, a process which becomes more frequent as the 
infection advances. The nucleus throws out a portion of its 
chromatin, which moves outwards, but remains connected with it by 
means of a thick band. As this chromatic mass approaches the 
periphery of the parasite, the cytoplasm bulges out from the surface 
and concentrates itself around the terminal enlargement of the 
chromatic band. The latter structure becomes thinner and finally 
breaks. The connection of the bud with the main mass becomes in 
the meanwhile less and less extensive, and finally the bud is detached 
(figs. 10-12). Very often two buds are formed at the same time in a 
similar manner (figs. 13-16). A large number of buds as described by 
Kinoshita 7 have never been distinctly observed by us. 

Schaudinn 21 and Liihe 13 were the first to draw attention to the 
presence of a small nucleus in Piroplasma . This discovery has been 
confirmed by different workers. Schaudinn named the second nucleus 
a blepharoplast, and most of the later workers adhere to this view, 
without, however, producing any evidence in support of it. 

Our observations show that but few binucleate parasites are 
present at an early stage of the infection. This small nucleus then 
arises from the large one, usually at a later stage of the disease. 
Different phases of this process may sometimes be seen in one red 
corpuscle containing several parasites (figs. 17, 18, 42). The nucleus, 
which at this stage is surrounded by a vacuole, buds off a small part 
of its substance, which moves to the edge of the vacuole, often leaving 
a thin connecting line behind. 



236 


LATER FORMS OF PARASITES IN THE BLOOD 

As the infection advances, the parasites undergo marked changes, 
and only now and again large amoeboid forms are seen. The 
parasites diminish in size, and are frequently pear-shaped. The proto¬ 
plasm, which at first is a typical ‘ Schaumplasma,’ becomes much 
denser in structure. The percentage of binucleate forms increases, 
and many free forms are encountered. 

A peculiar feature of this stage is the detachment of small parts 
of the cytoplasm in a definite way. At one side of the cell appears 
a vacuole, which increases in size and enlarges within the parasite, 
until the protoplasm is almost separated into two unequal parts, which 
finally become separate. The smaller part is entirely cytoplasmic 
in nature (figs. 22-26). 

Owing to the rapidity with which multiplication takes place, the 
nuclear details become very irregular, and frequently a second division 
commences before the completion of the first (figs. 2o, 51-52). 

The nucleus of the round forms is usually surrounded by a 
vacuole (fig. 33). The division is by simple fission, in which the 
nucleus divides with the vacuole (figs. 33-38). 

Sometimes the parasites assume a signet-ring form, a large vacuole 
occupying the middle of the cell, the nucleus which lies at the 
periphery often dividing (fig. 21). 

The usual mode of division at this stage results in the formation 
of two pear-shaped forms, but differs from that in the early stages of 
the disease. Starting again from the round binucleate form, either 
the large or the small nucleus divides, together with the line; the 
small nucleus moves to the edge of the parasite, and the chromatic 
line becomes fainter, and in many instances finally disappears (figs. 
45-52). The divided large nuclei frequently remain connected 
(fig. 46). At this stage one or two vacuoles appear about the middle 
of the cell, and increase in size. Often the two parasites are 
connected by three fine protoplasmic strands, two peripheral and one 
across the middle, a large and small nucleus in each half. First the 
central connecting strand breaks, and the separation of the two 
parasites becomes more pronounced, until they are only connected by 
one strand at their apices. Whilst the pear-shaped forms are still 
connected in the above described manner, the connecting cytoplasmic 



237 


strand may be seen considerably thickened at the middle (fig. 19). 
This connection becomes smaller and smaller until both parasites 
separate into two pear-shaped forms; the small nucleus dividing again 
even before the separation is complete. On the other hand division 
of the large nucleus may set in first, accompanied or not, by a division 
of the connecting line (figs. 44-52). 

While the pear-shaped forms are still connected, a second small 
nucleus may arise from the large one (fig. 53 « 2 ). We have not been 
able to explain the meaning of this process. 

A striking feature of the present stage of the infection is the 
occurrence of unequal divisions of the parasite. The nuclei of the 
cells divide in the usual way, but the cytoplasm divides into two 
unequal parts, the smaller parasite assuming a crescent shape. This 
division may be compared with the sickle-shaped detachment of the 
cytoplasm described above. 

Leishman and Statham 11 describe a similar process in Leishmania 
donovani {Piroplasma dortovani ), with the important difference, 
however, that eventually nuclei were seen in these detached parts of 
the parasite. 

We, however, could not follow an analogous procedure in 
Piroplasma canis To our minds there are two distinct processes. 
Either the cytoplasm becomes detached in a regular way without 
co-operation of the nuclei; or, the nuclei take part in the division. 
The enucleated particles of cytoplasm probably degenerate, and give 
rise to the appearance of irregular dark staining masses in the 
protoplasm of the infected red cells, or the detached part contains one 
or two nuclei and gives rise to a new parasite (figs. 28 and 29). Rarely 
chromatin appears to be given off from the nucleus, and become free 
in the red cell (fig. 30). This process has already been described by 
Nuttall and Graham-Smith, 18 but its significance is unknown. 

The parasites occurring in the blood of organs do not differ 
markedly from those found in the peripheral and heart blood. As 
division appears to proceed more rapidly in the organ blood, the 
parasites are usually slightly smaller and more compact. The free 
forms, which occur in greater numbers in the organs, divide in the 
same way as the intra-cellular forms, i.e., round and pear-shaped 
division. (Compare fig. 43.) 



FLAGELLATED FORMS 


Flagella-like processes in different species of Piroplasma in blood 
have been frequently described. Bowhill and Le Doux, 2 Nuttall and 
Graham-Smith, 18 and Kinoshita, 7 describe their occurrence in 
Piroplasma cams ; Lignieres 12 and Bowhill 1 in cattle piroplasmosis ; 
Fantham 5 in Piroplasma muris. These processes have been more 
frequently observed in cultivation forms, and in developmental forms 
in the tick, by Koch, 9 Kleine, 8 Kinoshita, 7 and Miyajima. 14 

The meaning of some of these forms has been explained in 
different ways. Doflein, 4 Nuttall and Graham-Smith, 17 and 
Hartmann 6 discuss the probability of their being mikrogametes 
analogous to the mikrogametes of the life cycle in malaria, but nothing 
in the nature of a proof of this conception has hitherto been brought 
forward. When we consider the active amoeboid movement of the 
young parasites, it would certainly appear that most of the flagella¬ 
like processes seen must be regarded simply as fine pseudopodia. 
Kinoshita, 7 on the other hand (figs. 41 and 46), figures a flagellum 
which arises from a blepharoplast and takes a chromatic stain in the 
same way as do trypanosome flagella. 

Now and again, long flagella-like processes, which were evidently 
pseudopodia, have been seen in intra-corpuscular forms (figs. 31, 32). 
(Compare Kinoshita, fig. 9.) 

Very rarely true small flagellate forms were seen, especially in 
blood from the lung; but we were never able to trace the origin of 
the single flagellum (fig. 27). 

Large flagellated forms have been described by Miyajima 14 in 
cultures of Piroplasma parvum , and these forms he describes as 
intermediate stages in the development of trypanosomes from a 
typical Piroplasma. He discusses at length the possibility of a 
mixed infection of piroplasmosis and trypanosomiasis of the blood 
used for his culture experiments, but the facts he brings forward seem 
to be very much against such a possibility. 

Kossel’s and Weber’s observation, as quoted by Schaudinn, 21 seems 
to have anticipated Miyajima’s observation with regard to large 
flagellated forms, with the difference that they observed his culture 
forms in freshly drawn blood. 

Nuttall and Graham-Smith 17 in 1905 were the first to describe a 



239 


few large forms of Piroplasma canis , which simulate the crescents of 
aestivo-autumnal malaria, having the chromatin sometimes concen¬ 
trated in the middle, sometimes forming a loose mesh work. These 
parasites were 10*4 to 107 fi long and 1*4 to 17 fi broad. They 
regarded them at first as gametes, but in their last paper they do not 
consider them to have any connection with Piroplasma canis. Their 
chief reason was the fact that they only found seven ‘ gametoid 
bodies * altogether, and these occurred in one animal. No flagella 
could be observed. 

Kinoshita figures somewhat similar parasites (figs. 47, 48, 49) seen 
in the heart-blood, pancreas and lung, some hours after death. He 
refers to his figs. 47 and 48 as conjugation forms, and to fig. 49 as 
an ookinet (?) in accordance with Schaudinn’s ideas. 

We have been able to trace the development of large biflagellate 
forms from the normal intra-cellular parasite. In the films, where 
large biflagellate forms occurred, along with ordinary intra-cellular 
parasites, forms were also found in which both nuclei were 
considerably enlarged, as represented in figs. 54, 55. These bodies 
increase in size, and the smaller nucleus in the meanwhile divides 
(fig. 56), often remaining connected with the large one by fine 
chromatic lines. The subsequent changes vary in details, but on the 
whole two main forms of development may be followed. On the one 
hand, an irregular number of small round chromatin masses originate 
from the large nucleus, frequently remaining connected with it by fine 
chromatic lines, which eventually disappear (figs. 72, 73). From the 
fact that these masses often appear double, it seems possible that they 
may divide (figs. 75, 76). At the same time the appearance of the 
large nucleus changes; the chromatin becomes aggregated at the 
centre, and a lightly staining area is left between it and the well- 
defined nuclear membrane (figs. 60, 72-74). Eventually, two flagella 
are formed, each of which may end in the neighbourhood of a small 
chromatic mass, but in some cases the flagella appear to have no 
definite origin (figs. 74, 77, 78). 

A second mode of development takes a somewhat different line. 
The large nucleus frequently buds off at first a small number of 
granules, and eventually it seems to throw out the whole of its 
chromatin in form of a large densely staining mass; figures 62-66 
representing different stages of this process. The remainder of the 



240 


original nucleus persists as a homogeneous lightly staining mass, 
which retains its original form and moves to one side of the parasite. 
At this stage usually one long flagellum arises in fhe neighbourhood 
of a small chromatic body very often situated at one end of the 
parasite (figs. 61-64). Shortly after, a second flagellum is formed, 
sometimes arising in close vicinity to the origin of the first flagellum, 
sometimes at some distance (fig. 65). 

The above described development is very liable to modifications. 
Occasionally two large masses of chromatin are thrown out of the 
nucleus (fig. 68), and at the same time these latter sometimes divide 

( fi g- 70 - 

Whilst these nuclear changes are taking place, the parasites 
increase in size, and become elongated. The protoplasm changes its 
appearance, and becomes very loose and vacuolated. The dimensions 
of fully developed flagellate forms vary between 6 to 8 in length 
and 2 to 3 fi in width. 

These forms have been repeatedly observed by us in very small 
numbers in the peripheral blood of dogs on the day before death. 
Only in one animal were they abundant, and only in this case have 
we been able to follow their development. The blood was taken in 
the morning of the day before death. Films made actually on the 
day of death did not show any of these forms, neither in the peripheral 
nor in the heart blood, only two of these flagellated cells being found 
in organ films (spleen and bone-marrow). 

This observation seems to point to the fact that the biflagellate 
forms of Piroplasma canis represent a very transient stage in its life- 
history. For this reason, it might have been very easily overlooked. 
We, however, at present, are not able to form a definite opinion as to 
the significance of this stage in the life-history of the parasite, 
especially as the subsequent development of the flagellate forms could 
not be traced. 

Up to the present, no observations, either in culture or in the 
intermediate host, throw any light upon their meaning. Develop¬ 
mental stages of these flagellate forms in some respects resemble 
those occurring in the development of the flagellate forms in the 
cultures of Leishmania donovani. 



241 


LITERATURE 

1. Bowhill, 1903. Practical observations on Texas fever. Vet. Jour. Vol. XXXI. 

2. Bowhill and le Doux, 1904. A contribution to the study of Piroplasma 

cants. Journal of Hygiene, Vol IV. 

3. Christophers, 1907. Piroflasma cants and its life-cycle in the Tick. 

Scientific memoirs by Officers of the Med. and Sanit. Dpt. of the Govern¬ 
ment of India, No. 29. 

4. Doflein, 1901. Die Protozoen als Krankheitserreger. Fischer, Jena. 

5. Fantham, 1906. Piroflasma muris. Quart. Jour. Mic. Sci., Vol. L. 

6. Hartmann, 1907. Archiv. f. Protistenkunde, Vol. X. 

7. Kinoshita, 1907. Untersuchungen fiber Babesia cants. Arch. f. 

Protistenkunde, Bd. VIII. Heft. 2/3. 

8. Kleine, 1906. Kultivierungsversuche der Hundepiroplasmen. Zeitschrift f. 

Hyg. und Infektionskrankheiten, Bd. 54. 

9. Koch, 1905. Vorlaufige Mittheilungen iiber die Ergebnisse einer 

Forschungsreise nach Ostafrika. Deutsche med. Wschft., No. 47. 

10. Koch, 1906. Beitrage zur Entwicklungsgeschichte der Piroplasraen. 

Zeitschrift f. Hyg. und Infektionskrankheiten, Bd. 54. 

11. Leishman and Statham, 1905. The development of the Leishman bodies 

in cultivation. Journal of the Royal Army medical corps, Vol. IV. 

12. Ligni£res, 1903. La piroplasmose bovine. Arch, de Parasitologie, Vol. VII. 

13. Luhe, 1906. In Mense’s Handbuch der Tropenkrankheiten, Vol III. 

14. Miyajima, 1907. On the cultivation of a bovine Piroplasma. The Philippine 

Journal of Science, Vol. II, No. 2. 

15. Moore and Breinl, 1907. The cytology of Trypanosomes, Part I. Annals 

of Trop. Med. and Parasitology, Vol. 1, No. 3, Appendix I, II. 

16. Nuttall, 1904. Canine Piroplasmosis I. Jour, of Hygiene IV. 

17. Nuttall and Graham-Smith, 1906. Canine Piroplasmosis II. Jour, of 

Hygiene V. 

18. Nuttall and Graham-Smith, 1906. Canine Piroplasmosis V. Jour, of 

Hygiene VI. 

19. Nuttall and Graham-Smith, 1907. Canine Piroplasmosis VI. Jour, oi 

Hygiene VII. 

20. Piana-Galli Valerio. Moderno Zoojatro, 1895. 

21. Schaudinn, 1904. Generations und Wirtswechsel bei Trypanosoma und 

Spirochaete. Arb. a. d. Kaiserl. Gesundheitsamte, Vol. XX. 



242 


EXPLANATION OF THE PLATES 

All the figures are drawn with a Zeiss apochromatic 2 mm. oil 
immersion lens, aperture 1*40. Oc. 18. 

Plate VI 

Figs. 1-16.—Breinl’s stain. 

Fig. 1.—Early amoeboid form. 

Figs. 2-4.—Division stages of amoeboid forms. 

Fig. 5.—Binucleate amoeboid form. 

Figs. 6-9.—Pear-shaped division of binucleate form. 

Figs. 10-12.—Stages in the formation of a single bud. 

Figs. 13-16. Stages in the formation of two buds. 



Breird <V HindLe 


Plate VI. 





R 



2 44 


Plate VII 


Figs. 17-27.—BreinTs stain. 

Figs. 28-32.—Heidenhain-Breinrs stain- 

Figs. 17, 18.—Different stages of the formation of the small nucleus. 

Fig. 19.—Pear-shaped division form, showing thickening of the 
connecting line. 

Fig. 20.—Pear-shaped division; division of small nuclei before 
separation. 

Fig. 21.—Signet ring form, with divided nucleus. 

Figs. 22-26.—Formation of the sickle-shaped mass of cytoplasm. 

Fig. 22.—Appearance of vacuole at the edge of the cell. 

Figs. 23, 24.—Growth of the vacuole. 

Figs. 25, 26—Separation of the sickle-shaped cytoplasmic part. 

Fig. 27.—( a ) Free pear-shaped binuclear form. 

( b ) Free small flagellate parasite. 

Figs. 28, 29.—Unequal division. 

Fig. 30.—Extrusion of chromatin into the red cell. 

Fig- 3 1 -—Amoeboid form with long pseudopodium. 

Fig- 32.—Division of same. 



Bread. r> Hindle. Plate ITT. 

17 . 18 . 19 . 20 . 



Kuth, LI L o d o: i. 


I '.male a.d na.t df.l 







246 


Plate VIII 

Figs. 35-48.—Heidenhain’s stain. Later stages of division. 

Figs. 33-40.—Division of round forms. 

Fig. 41.—Round intra-cellular binucleate form. 

Fig. 42.—Formation of small nucleus. 

Fig. 43.— (a-c) Division of free round forms, (d) Formation of small 
nucleus in free form. 

Figs. 44-48.—Stages in the late pear-shaped division. 

Fig. 44.—Division of large nucleus and chromatic line 

Fig. 45. In the left parasite division of small nucleus. In right 
parasite appearance of vacuole and commencement of 
division of large nucleus. 

Fig. 46.—Both nuclei divided; daughter cells connected by three 
strands. 

Fig. 47.—Disappearance of middle connecting strand. 

Fig. 48. -Rupture of lower connecting strand. 








248 


Plate IX 

Figs. 49-78.—Heidenhain-Breinl’s stain. 

Figs. 49-50.—End stages of pear-shaped division. 

Fig. 51.—Division of small nuclei before separation of daughter cells. 
Fig. 52.—Division of large nuclei before separation. 

Fig. 53.—Formation of a second small nucleus (ft). 

Figs. 54-78.—Formation of large biflagellate parasites. 

Figs. 54, 55.—Swelling up of nuclei, in intra-cellular parasites. 

Fig. 56.—Division of enlarged small nucleus. 

Fig- 57-—Extrusion of chromatin from the nucleus. 

Figs. 58, 59.—Swelling up of nucleus in three parasites. 

Fig. 60.—Transformation of large nucleus. 

Fig. 61.—Formation of flagella. 

Figs. 62-64.—Stages in the extrusion of chromatin from the nucleus. 
Fig. 65.—Formation of second flagellum. 

Figs. 66, 67 and 69.—Large elongated biflagellate forms. 

Fig. 68.—Extrusion of two masses of chromatin from the nucleus. 

Fig. 70.—Large flagellate form after extrusion of chromatin, 
containing a number of large granules. 

Fig. 71.—Division of extruded chromatin and disappearance of the 
remains of the original nucleus. 

Figs. 72-78.—Stages in the development of large biflagellate forms 
with characteristic nuclei. 







249 


COMPARATIVE 

CHEMO-THERAPEUTICAL STUDY OF 
ATOXYL AND TRYPANOCIDES 

PART I 

BY 

M. NIERENSTEIN, Ph.D. 

J. W. GARRETT INTERNATIONAL FELLOW AND RESEARCH DEMONSTRATOR OF THE 
LIVERPOOL SCHOOL OF TROPICAL MEDICINE 

(From the Runcorn Research Laboratories) 

(Received for publication 14 May, 1908) 

Atoxyl , sodium-p-amino phenyl-arsenate was introduced in the 
treatment of Trypanosomiasis by Thomas and Breinl 1 (1905), and its 
specific therapeutic value for sleeping sickness has been more or less 
recognised. It contains from 25*95 to 20*78 per cent, of arsenic; the 
difference depends on the water of crystallisation, as shown by Moore, 
Nierenstein and Todd, 2 Ehrlich and Bertheim, 3 and others 

Arsenic in the form of Atoxyl is much better tolerated by the 
animal organism than in the form of sodium arsenate ; the therapeutic 
value of the Atoxyl, therefore, was attributed to the fact that much 
more arsenic could be administered in this new form. It was supposed 
to act simply as an internal antiseptic , and was thought to kill the 
parasites in direct proportion to the amount of arsenic introduced. 

Some experiments made in June, 1907, by Breinl and Nierenstein 
seemed to disprove this idea. In an attempt to produce an active 
immunity against Ngana, mixtures of Atoxyl and trypanosomes were 
injected in different proportions, and after different periods of contact, 
with the idea that by increasing the amount of trypanosome-infected 
blood and decreasing the amount of Atoxyl, and by lessening the time 
of contact, a point might be reached at which virulent trypanosomes 
could be injected with impunity. 



250 


The results obtained, however, were not what were expected ; 
dogs, rabbits and donkeys were used for the experiments, but 
invariably after the first injection, even after exposure of the mixture 
for forty-five minutes to a temperature of 37 0 C., the animals became 
infected after a normal incubation period. This fact seemed to 
suggest that the action of Atoxyl was not simply disinfectant, but was 
the result of a co-operation between the living tissues and the drug. 

Uhlenhuth, Hiibner and Woithe 4 in their experimental study of 
the action of Atoxyl on T. cquiperdum came to a similar conclusion. 
They state (p. 296) : — 

4 Unsere Meinung gelit jedenfalls dahin, dass der Chemismus der 
Atoxylwirkung kein so einfacher ist, wie ihn die Theorie der 
Arsenspaltung supponiert, das vielmehr beim Zustandekommen des 
wunderbaren therapeutischen Effektes die Korperzelle eine ganz 
hervorragende Rolle spielt.’ 

This observation of ours, confirmed by Uhlenhuth, Hiibner and 
Woithe, was the starting point for the following study of the 
therapeutical action of Atoxyl. 

The experiments were divided into two groups, the action of 
Atoxyl and similar compounds on serum proteids being studied 
in vitro and in vivo , respectively. Only the results of the first series 
are here recorded 

Technique .—20c.c. of normal serum and 20C.C. of a 2 per cent, 
solution of the compound were shaken up for twenty-four hours, and 
the proteids afterwards were precipitated with 35 c.c. of a 2 per cent, 
solution of tannic acid. The precipitate was then carefully washed 
for about forty-eight hours and arsenic estimations of the filtrate were 
made from time to time until no trace of arsenic could be found in the 
filtrate. The precipitate was treated with 10-15C.C. of concentrated 
sulphuric acid, and digested in a Kjeldahl flask in the usual way. 

The arsenic estimations were made by Sanger’s 5 method. Instead 
of hydrochloric acid, gold chloride was used as a developer, and proved 
much more sensitive. 

In those cases in which arsenic was found in the precipitate after 
digestion, some of the original product was dialysed against water in 
a parchment sausage-skin, and the dialysate was evaporated to 
dryness and tested for arsenic. 



251 


i. Atoxyl and Serum 




A *V°N. 

/V° 

Atoxyl = | ! 

\/ 

NH, 



Srrtxm Used. 

Arsenic Estimation 
in Precipitate. 

Arsenic Estimation 
IN Dl A I. VS ATE. 

Horse serum 

arsenic present 

- 

- 

Horse serum 

arsenic present 

arsenic 

absent 

Horse serum 

arsenic present 

arsenic 

absent 

Goat serum 

arsenic present 

arsenic 

absent 

Goat serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 

Donkey 

serum 

arsenic present 

arsenic 

absent 


2 . 

Sodium Arsenate and 

Serum 

Serum Used. 

Arsenic Estimation 
in Precipitate. 

Arsenic Estimation 
in Diai.ysatk. 

Horse serum 

arsenic absent 

— 

Horse serum 

arsenic absent 

— 

Horse serum 

arsenic absent 

— 

Goat serum 

arsenic absent 


Goat serum 

arsenic present 

arsenic present 

Goat serum 

arsenic absent 

-- 

Goat serum 

arsenic absent 

- - 

Donkey serum 

arsenic absent 

— 

Donkey serum 

arsenic absent 

— 

Donkey serum 

arsenic absent 

— 

Donkey serum 

arsenic absent 

— 

Donkey serum 

arsenic absent 

— 



3. Acetylated Atoxyl and Serum. 





Mono-acetylated atoxyl = | | 

V > 



n/OCH, 


Serum Used. 

Arsenic Estimation 

Arsenic Estimation 

in Precipitate. 

in Diai/ysate. 

Donkey serum 

arsenic present 

arsenic absent 

Donkey serum 

arsenic present 

arsenic absent 

Donkey serum 

arsenic present 

arsenic absent 

Donkey serum 

arsenic present 

arsenic absent ( ?) 

Donkey serum 

arsenic present 

arsenic absent 

4. Benzoylated Atoxyl and 

Serum 


AsO/ ONa 
As °\OH 


Mono-benzoylated atoxyl = 




Serum Used. 


Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 


^/CO.C # H 

'H 

Arsenic Estimation 
in Precipitate. 

arsenic present 
arsenic present 
arsenic present 
arsenic present 
arsenic present 


Arsenic Estimation 
in Dialysate. 

arsenic absent 
arsenic absent 
arsenic absent 
arsenic absent 
arsenic absent 


5. Benzoyl-acetyl-atoxyl and Serum 


Benzoyl-acetyl-atoxyl 


AsO/ONa 

0 

N/CO.CH, 

SCO.C.H 


6 


Serum Used. 


Arsenic Estimation 
in Precipitate. 


Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 


arsenic absent 
arsenic absent 
arsenic absent 
arsenic absent 
arsenic absent 



253 


6. SODIUM-p-HYDROXYL-PHENYL-ARSENATE* AND SERUM 

Sodium-p-hydroxyl-phenyl- 

arsenate = 

OH 

Arsenic Estimation- 
in Precipitate. 

arsenic absent 

arsenic absent 

arsenic absent 

arsenic absent 

arsenic absent 

arsenic absent 

arsenic absent 

It is evident from the foregoing experiments that a combination 
takes place respectively between proteins and Atoxyl, mono- 
acetylated Atoxyl, and mono-benzoylated Atoxyl; whilst no 
combination occurs respectively between these proteins and sodium 
arsenate, acetyl-benzoyl Atoxyl, and sodium-p-hydroxy-phenyl- 
arsenate. 

It might be mentioned that there is a considerable difference in 
the results obtained by the treatment of Trypanosomiasis by means 
of the above-mentioned compounds. Whereas Atoxyl and mono- 
acetylated Atoxyl act promptly on the parasites, the effect of sodium 
arsenate is less pronounced, that of sodium-p-hydroxy-phenyl-arsenate 
is nil. 

The analogy between the way in which these compounds behave 
with proteins, and their action on trypanosomes, is very suggestive. 
We are, hence, led to believe that this combination with the proteins 
is of importance in trypanocidal drugs, and have now to consider how 
Atoxyl and its derivatives become attached to the proteins. 

Ehrlich 6 has compared the action of a drug to that of a dye. We 
know that it is necessary for a dye to possess a chromophoric group— 
a chemical radical which causes it to be coloured—and a chromogenic 

• Our thanks are due to Messrs. Burroughs, Wellcome & Co., who kindly supplied 
this drug. 


Serum Used. 

Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 
Donkey serum 





2 54 


group, which renders it a dye. This is easily illustrated by the 
following example: — 

Azo-benzene (C 6 H 5 N = NC 6 H 6 ), which contains the chromophor 
N — N, is coloured, but does not possess dyeing properties. It only 
becomes a dye when the chromogenic group OH or NH 2 enters. 
Similarly, for example, oxyazo-benzene (OH.C e H 4 N — NC 6 H 5 ) and 
amino-azobenzene (H a N.C 6 H 4 N = N^Hg) are dyes. Their dyeing 
value increases with the number of chromogenic groups introduced. 
For this reason tri-amino-benzene (NH 2 .C fl FI 5 N = N.C 6 H 3 (NH 2 ) 2 ) is 
a much better dye than amino-azo-benzene. 

When wc apply the same theory to the therapeutics of Atoxyl, we 

y yONax 

find that sodium-phenyl-arsenate ( CeH 5 AsO\ ) (which has 

V \oh y 

been proved by Plimmer and Thomson 7 , and also in this labora¬ 

tory, not to possess any curative effect), and also sodium-p-hydroxy- 

y yONax 

phenvl-arsenate ( OH.CgH 4 AsO\ ) do not combine with the 

V \OH y 

y yONaX 

proteins, whilst atoxyl ( NH 2 CgH4AsO\ ) combines with the 

V xoh y 


combines with the 


proteins and acts on trypanosomes; mono-acetylated atoxyl 

y yONax 

( CH a CONH.C6H 5 AsO\ Jcombines and is curative, while fully 

V \oh y 

y CH 8 COx ^ONax 

acetylated and benzoylated atoxyll yN.CeHgAsO^ ) 

XCeHsCOy NOH / 

does neither. 


Hence, we suggest that in Atoxyl the amido group (NH 2 -group) 
and in mono-acetylated Atoxyl the imido group (NH-group) play the 
same role as the chromogenic group in a dye. It has already been 
pointed out that the action of Atoxyl has generally been explained as 
being due to the arsenic, and the advantage of its use is that more 
arsenic could be introduced in the organism in form of Atoxyl than 
in form of sodium arsenate; it might be argued from this point of 
view that the action of Atoxyl is as follows: — 

The Atoxyl attaches itself to the proteins; the benzene nucleus is 
slowly oxydised by the tissues and the arsenic is set free; so that, 



2 55 

when combined with the tissues, Atoxyl acts as a storage for effective 
arsenic. 

This, however, is apparently not the case. It is well known that 
Trypanred, Afridol blue and Afridol violet, also Parafuchsin, have an 
effect on trypanosomes comparable to that of Atoxyl. These 
compounds do not contain arsenic, but a large number of amido 
groups. Further, Laveran, 8 also Thomas and Breinl, have found that 
sodium arsenate in combination with trypanred acts much better than 
sodium arsenate alone. 

We have, therefore, reason to believe that the amido group in 
Atoxyl, and in the above-mentioned colouring matters, has a specific 
action on trypanosomes, and that in Atoxyl the effective part is not 
only the arsenic, but also the amido group. 

How this group acts on the parasites is engaging our attention at 
present, and will form the subject of a subsequent communication. 


LITERATURE 

1. Thomas and Breinl. Liverpool School of Tropical Medicine, Memoir XVI, u>o6. 

2. Moore, Nierenstein and Toon. Bio-Chemical Journal, Vol. II, p. 300, 1907. 

3. Ehrlich and Bertheim. Ber. d. deutschen chem. Gesellschaft, Vol. XL, p. 3202, 

(19<>7)- 

4. UHLENHUTH, Hubnkr and Woithe. Arbeiten aus den Kaiserlichen Gesundheits- 

amte, p. 256, 1907. 

5. Sanger and Black. Jour. Soc. Chem. Inst., Vol. XXVI, p. 1115, 1907. 

U . Ehrlich. Deutsche med. Wochenschrift, p. 1052, 19 >8 ; also Festschrift fiir Leyden, 
Vol. I, p. 645, and Internationale Beitrage zur innern Medizin, Berlin, 1902. 

7. Plimmkr and Thomson. Proceedings Royal Society, Vol. LXXIX, pp. 505-16 

(1907). 

8. Laveran. C. R. de TAcad. des Sciences, Vol. CXL, p. 108, 17 April, 1905. 




2 57 


ON THREE NEW SPECIES OF CULEX COL¬ 
LECTED DURING THE ANTI-MALARIAL 
CAMPAIGN IN MAURITIUS IN 1908* 


BY 

d’EMMEREZ de CHARMOY 

(WITH ONE PLATE) 

(Received for publication 12 April , 1908) 

Culex arboricollis , n. sp. 

Male. — Head: eyes greenish ; occiput yellowish, with long, white 
and yellow, curved scales and a few hair-like black scales; the 
yellow scales are placed closely in the line separating the eyes. The 
antennae bear long hairs which are pale yellowish apically and 
greyish-black basally; the segments of the basal half furnished with 
very long, narrow curved scales; t apical segment with a few short 
hairs ; the basal segment with short, flat, white scales. Palpi of four 
segments, as long as the proboscis, with narrow white bands at the 
base and the apex of the second, third and fourth segments; white 
scales are desseminated over all the segments. Proboscis black, with 
the apex paler and a yellowish band in the middle. 

Thorax black, covered sparsely with long, narrow curved, white 
and golden scales, and long, black, hair-like scales; those portions 
of the thorax which are not covered with scales form velvet black 
spots. Scutellum bordered with flat whitish scales, and dark hair¬ 
like scales ; metanotum nude, black. 

Abdomen velvety black, with whitish basal bands ; apical segment 
with a few whitish scales at apex; all the segments with long 
yellowish marginal hairs. 

Legs black, with more or less loosely scattered, yellowish scales ; 
the articulations of the femora and tibiae are basally and apically 
banded; the tarsi are black without any coloured scales; metatarsi 
of front legs are basally banded ; the other tarsal segments are black ; 
in the mid leg the metatarsi and the first tarsal segments are basally 

* During the Expedition of Professor Ross, F.R.S., C.B. 

f This remarkable insect bears some resemblance to Theobald’s genus Lophoceratoinyia , 
though it is quite distinct, and a new genus will probably be erected to receive it.— 
R. Newstead. 


banded; in the hind legs the metatarsi are basally banded with 
yellowish scales, and apically with white scales; all the remaining 
segments are basally and apically banded with white scales. 

Wings spotted. The black spots on the costa extend to the 
auxiliary vein. They are seven in number and are situated as 
follows: two small basal ones, the second a little larger than the 
first, the third one having a white dot in its middle, the fourth and 
fifth ones united on the auxiliary vein by black scales, the sixth 
placed obliquely, the seventh near the apex; the other veins are 
irregularly spotted with white scales, the last vein* which bears the 
black fringe is regularly spotted white and black on its basal half. 
The underside of the body presents the following markings: The 
pleura densely covered with imbricated, flat, whitish scales; the 
trochantae, coxae, and the base of the femora are covered with white 
scales; the ventral segments of the abdomen are spotted basally with 
white scales and apically with a well-defined, narrow, white line. 

FEMALE. —Proboscis black, with a few scattered white scales, and 
a white band just below the first anterior third. Palpi longer than 
the half of the proboscis, with a few scattered white scales and white 
bands; the apical segment bears two moderately long hairs. The 
fore part of the occiput is covered with long, narrow curved, white 
scales; the hind portion with yellow, upright forked scales; the 
anterior lateral portions with black, upright forked scales. Scutellum 
with a median and two lateral tufts of long, black hairs, and a few 
long, flat, curved white scales. The thorax and pleurae as in the 
male. Halteres yellowish, with small, white scales. The larvae of 
this species were found by Professor Ronald Ross in the holes of 
trees at Vacoas; and although the larval habitat was situate near 
dwellings, no adults were seen in houses or verandahs. This well- 
marked species is apparently uncommon, and comes near Culex 
mimeticus . 

Culex fowleri , n. sp. 

Female. — Proboscis brown, base paler, with whitish scales. 
Palpi black, with a few long, black hairs ; the apex white. Antennae 
brown, spotted with white; first segment bearing white scales. 
Occiput at the sides covered with flat, imbricated, white and black 


• ? Costa.—R. N. 



2 59 


scales; the median portion covered with long, white, narrow, curved 
scales, black, upright forked scales, and black hair-like scales. 

Thorax brown, with two sub-median greyish lines, with long 
narrow curved, golden scales and black hair-like scales. • Scutellum 
with white scales and golden, hair-like scales. 

Abdomen black, with white basal bands. First abdominal segment 
with a basal white dot and apical white line, the other segments with 
apical white bands; the penultimate one with two apical spots; the 
last with lateral white spots. The underside of the abdomen with 
basal and apical bands. 

Legs . Under surface of the femora and trochantae of the 
posterior legs white; the upper surface brown, with small scattered 
spots; femora of the fore legs with white scales and hairs at their 
apices. Metatarsi with the first and second segments white at the 
base; the femora and tibiae of the mid legs are marked with white 
at their basal and apical parts. Metatarsi with the first and second 
segments white at the base. The femora of the hind legs white 
apically. The veins of the wings are covered with brown and white 
scales. Male similar. 

This species is easily distinguished from all other members of the 
genus Culex by the black and white spots oil the body of the insect. 
It was discovered by Major P. Fowler at Vacoa. 

Culex ronaldi , n. sp. 

Female.- Proboscis brown, with a yellowish median band. 
Palpi brown, bearing long hairs. Antennae brown; auxiliary hairs 
black, longer than those situated on the segments which are whitish. 
Eyes black. Occiput bearing scattered, white, long, narrow curved 
scales which form a continuous white line round the eyes. Upright 
forked scales black; hair-like scales black. Thorax brown, covered 
with long, narrow curved, golden scales, and long, black, hair-like 
scales, these are numerous on the posterior lateral margins. Scutellum 
with long, narrow, golden scales and black hair-like scales. Halteres 
white, yellowish at the tips. Abdomen black, with basal white bands. 
Scales of the wings brown. Legs : The femora are white apically; 
the tibiae white basally. Tarsi with narrow dusky white basal bands. 
Under side of trochantae and femora covered with white scales, 
s 



26o 


MALE. —Palpi a little lunger than the proboscis, brownish and 
hairy at the apex, with a white band at the base of the second 
segment and in the middle. The articulation of the segments of 
the antennae are black, the remaining portions of the segments 
whitish. Eyes black. The lateral portions of the head white; 
median portion with long, narrow, curved, white scales; upright, 
forked and hair-like scales, black. The under side of the abdominal 
segments with large lateral white bands, the penultimate one 
descending obliquely to the lateral margins. 

Found in the larval stage by Major P. Fowler in the broad moat 
outside Fanfava Bastian, in December, 1907, and January, 1908. 
Ground marshy, water from few inches to one foot deep, with much 
coarse grass. The larvae occurred in association with numbers of 
P, costal is, 

LIST OF CULICIDAE OF MAURITIUS 

ANOPHELINAE (Anophelines) 

1. Pyretophorus costalis , Loew (1866). 

Anopheles costalis , Loew (1866). 

A . gambiae , Giles (1902). 

A, gracilis , Donitz (1902). 

This species has been proved to be the principal carrier of Malaria 
at Phoenix and Vacoa where they are most numerous. Daruty and 
d’Emmerez found it very common at Port Louis in 1900. In some 
places near the sea shore it is uncommon, for example at Rre. At 
Seche and Maheburgh very few have been found. 

2. Myzorhynchus mauritianus , d’Emmerez and Daruty (1900). 

Anopheles paludis var. similis , Theobald (1901). 

A. tenebrosus , Donitz (1902). 

Very common everywhere and especially at Curepipe, Vacoa and 
Phoenix. All the specimens caught in the open air at Phoenix, 
Vacoa, where malaria is prevalent, were found not to be infected. 

j. Nyssorhynchus maculipalpis (Giles). 

Anopheles maculipalpis , Giles (1902). 

Not common; a few specimens only were caught by Major Fowler 
at Iron Fanfaren in Port Louis. 



26 i 


CULICINAE 

4. Stcgomyia scuiellarisf (Walker) (1859). 

Culcx scutellaris, Walker. 

C. albopictus , Skuse. 

C. variegatus , Doleschard. 

Very common everywhere, certainly the most abundant species of 
the island; the larvae occurred in tins, leaves, holes in trees and 
in the Ananas Sauvages. 

5. Stcgomyia fasciata , Fabricius (1805). 

Culex fasciatus , Fabricius (1805). 

C. calopus , Meigen (1818). 

C. tacniatus , Wiedemann (1898). 

C. clcgatts , Ficalbi (1896). 

C. rass», Giles (1899). 

C. cxagitans , Walker (1856). 

C. konuoupi , Brull6 (1832). 

C. zonatipes , Walker. 

C. formosus, Walker (1848). 

C.frater , Robineau-Desvoidy (1887). 

C. cxcitans , Walker (1848). 

C. viridifrons , Walker (1848). 

C. inexorabilis , Walker. 

C. bancrofti , Skuse (1886). 

C. mosquito , Aribalzaga (1891). 

C. annulitarsis, Macquart (1848). 

C. impatabilis , Walker (i860). 

Very common near the sea shore, in Port Louis; but rather scarce 
in the high parts of the islands. 

6. fWtf# arboricollis , n. sp., d’Emmerez de Charmoy (1908). 

The larvae of this interesting species were found in the holes of 
trees at Vacoa. It is, however, very scarce. 

7. ronaldi , n. sp., d’Emmerez de Charmoy (1908). 

Not common, the larvae were found at Iron Fanfaron. The larvae 
can be easily differentiated from those of the other species of this 
Island by its very long siphon tube. 

f Theobald (Genera Insectonim, p. 19, 1905) ^ives Scutomyia notoscripta (Skuse) 
priority.—R.N. 



26 2 


8. Culex annulioris , Theobald (1901). 

Only one specimen of this species was taken by Colonel Peterkin, at 
V acoa. 

9. Culex fou'leri , n. sp., d’Emmerez de Charmoy (1908). 

Not common. A few specimens obtained from larvae caught by 
Major P. Fowler. 

10. Culex tigripes , d’Emmerez and Daruty (1900). 

Culex maculicrura , Theobald (1901). 

Very common, and one of the largest species known. The larvae 
are carnivorous and they also eat each other. 

11. Culex fatigans, Wiedemann. 

Culex anxifer , Coquerel (Bigot). 

The commonest of all the species. It is very numerous all over the 
island and very troublesome during the night. The larva are to be 
seen in all artificial collections of water. 

12. Culex , spec, incert (male). 

A single specimen, caught by Major Fowler agrees in some respect 
with C. annul tor is \ but it is evidently distinct, though not 
sufficiently well preserved to render identification possible. 


ANOPHELINES THAT TRANSMIT MALARIA 

Pyretophorus costal is 

228 examples were caught at Clairfond Marsh between February 4, 
1908, and February 20, 1908. 73 of these were examined, of which 

10 were infected (i.e. 13*7 per cent.). 

? Myzorhynchus mauritianus 

54 examples which were fed on blood containing crescents and other 
gametes gave one positive result. (Round pigmented cells ? dead 
zygotes eight days after the first meal.) 56 other examples caught 
wild were negative. 




2()4 


EXPLANATION OF PLATE X 
Culex tigripes 

Fig. i.—Right ventral half of the head of the larva. 

Fig. 2.—Anal segments of the larva with siphon tube. 

Fig. 3.—Labial plate of the larva. 

Culex ronaldi , n. sp. (page 259) 

Fig. 4.—Anal segments of the larva with siphon tube. 

Fig. 5.- Antenna of the larva. 

Culex arboricollis , n. sp. (page 257) 

Fig. 6.—Wing of the male. 

Fig. 7, a , and b. —Claws of the tarsi. 

Fig. 8.- Proboscis and antenna of the female. 

Culex fowl err, n. sp. (page 258) 

Fig. Q.—Portion of the wing shewing distribution of the scales. 

Fig. 10.—Wing scales. 

Nyssorhynchus maculipalpis 

Fig. 11. Right dorsal portion of the head of the larva. 

Fig. 12--Anal segments of the larva. 

Fig. 13.—Second anterior abdominal segment of the larva shewing 
palmate and marginal hairs. 

Fig. 14.- Right anterior portion of the thorax of the larva. 






o.' >S 


if fid-> xi e(. 







2(>5 

CONCERNING THE TREATMENT OF 
EXPERIMENTAL TRYPANOSOMIASIS* 

PART II 

BY 

BENJAMIN MOORE, M.A., D.Sc., 

JOHNSTON PROFESSOR OF BIO-CHEMISTRY 

M. NIERENSTEIN, Ph.D., 

J. W. GARRETT INTERNATIONAL FELLOW, AND RESEARCH DEMONSTRATOR, LIVERPOOL 
SCHOOL OF TROPICAL MEDICINE 
AND 

JOHN L. TODD, M.D., 

ASSOCIATE PROFESSOR OF PARASITOLOGY, MCGILL UNIVERSITY 

From the Runcorn Research Laboratories of the Liverpool School of 

Tropical Medicine 

{Received for publication 14 July , 1908) 


INDEX PAGE 

I. Introduction . 266 

II. Completion of Part I . 268 

III. Inefficacious substances . 269 

IV. Aniline colouring matters . 269 

V. The therapeutic values of Atoxyl and acetylated Atoxyl 

compared . 272 

A. Dogs. 

1. Treatment by Atoxyl alone . 272 

2. Treatment by acetylated Atoxyl alone . 273 

3. Treatment by acetylated Atoxyl and bichloride of 

Mercury . 273 

B. Guinea Pigs. 

1. Treatment by Atoxyl alone . 274 

2. Treatment by acetylated Atoxyl followed by 

bichloride of Mercury . 275 

C. Mice. 

VI. Treatment by Atoxyl followed by another drug . 276 

VII. Treatment by Atoxyl followed by bichloride of Mercury 277 

A. Rabbits. 

1. Controls, treated by Atoxyl alone . 277 

2. Treatment by Atoxyl followed by bichloride of 

Mercury . 277 

(a) Infections with Trypanosoma brucei . 277 

(£) Infections with Trypanosoma gambiense . 279 

B. Donkeys. 

1. Controls, treated by Atoxyl alone . 280 

2. Treatment by Atoxyl followed by bichloride of 

Mercury . 282 

{a) Symptomatic treatment . 282 

(£) Routine treatment .!. 284 

VIII. Treatment by Trypanroth followed by bichloride of 

Mercury . 285 

IX. Summary and conclusions . 286 


* Grants in aid of this work were received by us from the Royal Society, and 
by one of us (Nierenstein) from the British Medical Association. We gratefully 
acknowledge the assistance received from these bodies. 

Our thanks are also due to Professor Ehrlich for acetylated Atoxyl (Acetyl- 
para-amino-phenyl-arsenic acid, 8 and 11); and to Messrs. Bayer and Co., 
Elberfeld, Germany, for the various colouring matters on which we report in 
Section TV. 

T 
























266 


I. INTRODUCTION 

This paper is a continuation of the experimental work on the 
treatment of experimental trypanosomiasis already published by 
us. 1, 2 

The technique of the present series of experiments was the same 
as employed in our former work. Trypanosoma brucei was again 
used in all of our experiments, unless it is otherwise specified, 
because of its great virulence for experimental animals, which makes 
it possible for conclusions to be drawn more quickly and more 
certainly from work done with it than is the case with any other 
pathogenic trypanosome. 

The strain with which we worked killed untreated rats in from 
three to five or seven days. In each series of experiments described, 
animals of about equal body weight were used as far as possible. 
All inoculations were made subcutaneously with blood mixed with 
Sodium citrate solution in saline, and approximately equal quantities 
of blood were used in every inoculation of animals of the same species 
where results were required for the comparison of different drugs. 
The usual quantity of infecting blood inoculated varied from I c.cm. 
to 5 c.cm. for rats to from 2 c.cm. to 5 c.cm. for guinea pigs and as 
much as 10 c.cm. for donkeys. The strain which was used for 
infecting purposes was always kept going in untreated animals, and 
therefore there is no possibility of a resistance to drugs, acquired by 
the trypanosomes, having militated against the success of our 
experiments. 

Treatment was commenced in no instance before there were 
definite signs of a well-established infection, such as the constant 
presence of the parasites in the peripheral circulation of the infected 
animal. 12 It may be noted here that, however valuable they may be 
as an indication of the trypanocidal value of the drugs used, 
experiments, in which treatment is commenced on the first or second 
day of the appearance of the trypanosomes in the blood, and 
therefore, before the infection is well-established, are, for practical 
purposes, valueless. Because, to give but one reason, it would 
certainly, as a rule, be practically impossible to treat a naturally 
infected animal at such an early stage of the disease. We suspect 
that early infections are so much the more easily treated not only 



267 


because there are fewer parasites present, but also because 
developmental, resistant forms have not yet been produced; we also 
suspect that it is to such resistant forms that recurrences are often 
due. 2 * 9 

Control animals of the same species which remained untreated 
were inoculated at the same time as the treated animals in every 
instance where the therapeutic effect of any drug was tried; and in 
experiments with Trypanosoma gambiense , additional controls were 
inoculated from the experimental animal just before its treatment 
was commenced, to definitely prove for each experiment the virulence 
of the infecting parasite. 

The Atoxyl used came from the Charlottenburg firm which first 
manufactured the drug. Unless it is otherwise stated, it was used 
in a freshly-made five per cent, solution in water previously sterilized 
at ioo°. The Mercury bichloride was usually employed in a one 
per cent, solution in water. The poisonous dose of each substance 
for the species of animal employed was always ascertained as a 
preliminary step to experimentation, and the largest possible 
therapeutic dose was used in each instance. Save when otherwise 
stated, all drugs were given subcutaneously. 

The routine examination of the blood was made in fresh three- 
quarter inch square coverslip preparations of blood from the tail or 
ear according to the animal. The blood of important animals was 
centrifugalised whenever it seemed necessary, as, for example, when 
trypanosomes could not be found in the blood by the ordinary 
examination, although the temperature was elevated. As a rule, for 
the first ten days or fortnight succeeding completion of the treatment 
the animals were examined daily. As they lived longer the 
examinations became less frequent, until they were done approxi¬ 
mately weekly or, in the case of experiments made with Trypano¬ 
soma gambiense , twice weekly. The blood in the less successful 
experiments, for example, the donkeys, was examined daily during 
the whole time these animals were under observation. The blood 
of any animal evidently ill was, of course, immediately examined, and 
if trypanosomes were not seen, subinoculations were at once made. 
Subinoculations of considerable quantities of blood were made also 
at intervals from animals which had been apparently successfully 
treated. All such subinoculated animals were kept under observation 



and examined frequently for at least three months in the case of 
Trypanosoma brucei , longer, if Trypanosoma gambiense was in 
question, before being considered to be uninfected. Rats and mice 
were, as a rule, used for such subinoculations in preference to other 
animals, as indeed they were throughout our entire work. 

II. COMPLETION OF PART 1 1 

The three control rats, treated by Atoxyl alone, which were alive 
at the date of publication of the last paper, have all since died of 
trypanosomiasis at periods varying between one hundred and two 
hundred and twenty-six days after the cessation of treatment. 

Trypanosomes (Trypanosoma brucei) have never recurred in any 
of the rats from which they had disappeared through the use of 
Atoxyl followed by bichloride of Mercury. All of the rats, save one, 
have died of pneumonia or have been killed because of skin diseases. 
None of the animals subinoculated from them, either before or at 
death, have become infected. One rat lived five hundred and 
nineteen days after inoculation, and then died from a skin affection. 
Nearly all the others had lived well over two hundred days before 
they died or were killed. It therefore seems justifiable to conclude 
that the rats, mentioned in this series of experiments, from which 
the trypanosomes were absent when Part I of this investigation was 
published, were definitely cured of their infection by Trypanosoma 
brucei . 

All the rats treated by Atoxyl followed by Donovan’s solution died 
of pneumonia in from one hundred and thirteen to one hundred and 
fifty-one days after inoculation; three of them died on the one 
hundred and fifty-first day. Trypanosomes had reappeared in none 
of them, nor have they appeared in the animals subinoculated from 
them. It seems probable that these animals were also cured of their 
trypanosome infection. 

It was shown by the inoculation and normally following death of 
a series of rats so cured that no immunity was acquired by animals 
which had recovered from an infection by Trypanosoma brucei after 
treatment with Atoxyl followed by Mercury; 11 neither was immunity 
conferred upon the young of such animals, for a series of eight young 
rabbits, both of whose parents had been treated and recovered from 
a trypanosome infection, became infected and quickly died on 
inoculation. 



269 

III. INEFFICACIOUS SUBSTANCES 

Because of the marked action of quinine on other protozoa, it 
was thought that substances derived from it, or resembling it, might 
be trypanocidal. Since Atoxyl, an organic arsenic-containing 
compound, kills trypanosomes, other compounds of like nature were 
tried alone, or in combination with various drugs; various other 
substances were tried in the same way. None of them were of value. 
The following are the drugs and combinations of drugs tried without 
advantage on rats infected with Trypanosoma brucei :~ 

Quinine. 

Cinchonine. 

Cupreine. 

Quinine-cacodylate. 

Quinine-cacodylate followed by Sublimate. 

Quinine-cacodylate followed by Iron-cacodylate. 

Quinine-cacodylate followed by Iron chloride. 

Arrhenal. 

Potassium bichromate. 

IV. ANILINE COLOURING MATTERS 

Our work in this direction is still very incomplete; we give, 
however, what results we have obtained. We have adopted the 
following routine method of gaining an idea of the efficiency of an 
untried substance. A series of rats is inoculated; some are left 
untreated, others are given one full dose of the substance to be tried. 
The average difference in days between the date of death of the 
controls and of the treated animals is noted. Its value is indicated 
for each of the drugs mentioned in the following list (column 3) ; 
our usual virulent strain of Trypanosoma brucei was the infecting 
parasite in each case. 


Phenolphthalein 

Average of 6 rats 

0 days 

Acetylated Phenolphthalein 

Average of 9 rats 

2 3 days 

Acetylated Phenolphthalein 
in alkaline solution 

Average of 4 rats 

3 days 

Methylated Phenolphthalein 

Average of 2 rats 

0 days 

Fluorescein 

Average of 4 rats 

0 days 

Acetylated Fluorescein 

Average of 3 rats 

1 day 



270 


Methylated Fluorescein 

Average of 4 rats 

0 days 

Eosin 

Average of 4 rats 

1*5 days 

Erythrosine 

Average of 4 rats 

0 days 

Floxine 

Average of 3 rats 

2 days 

Bengal rose 

Average of 2 rats 

0 days 

Rhodamine 

Average of 3 rats 

5-5 da ys 

Diazotized Rhodamine 

Average of 4 rats 

5 days 

Afridol violet 

Average of 3 rats 

4 days 

Afridol blue 

Average of 6 rats 

5 days 

Trypan red 

Average of 5 rats 

6 days 

Acetylated Rhodamine 

Average of 2 rats 

5 days 

Azogreen 

Average of 2 rats 

15 days 

Fuchsin 

Average of 7 rats 

8-5 days 

New Green 

Average of 6 rats 

0 days 

Crystal Violet 

Average of 6 rats 

0 days 

Antipyrin 

Average of 6 rats 

2*5 days 

A few experiments were 

made with Parafuchsin, 3 given 


subcutaneously; they were far from satisfactory. 

Since the discovery of ‘Trypanroth/ I, by Ehrlich and Shiga, 4 
great attention has been given to the trypanocidal properties of 
different aniline dyes belonging either to the Diazo-group, the 
* Afridols * of Mesnil and Nicolle, 5 or to the Triphenylmethane group, 
the 1 Malachite Green/ II, of Wendelstadt and Fellmer. 6 


NaO. 


/N = N—/\_/ \—N = N\ 

■ "'\A^ 


s w sa 


NaO.S' 


II 



N(CH,), 


Cl 


(CH.), 


v 

Vn |0-{-Qn(ch,) 



271 


Mesnil and Nicolle, in their excellent work on the action of 
Benzidines 7 (Diazo-colouring matters), have already pointed out that 
even the slightest difference in the constitution of a substance may 
have a great effect upon its trypanocidal action. This point is 
particularly well illustrated in the two following compounds, of which 
III has a very distinct effect upon the trypanosomes, while IV is 
practically non-trypanocidal. 



Our work upon the aniline colouring matters was undertaken 
with the intention of starting with an indifferent organic compound, 
that is, one which has no effect upon the trypanosomes, and of 
combining with it various radicles in the hope of finding one which 
would have definite trypanocidal properties. 

Our work has led us to the conclusion that the NH 2 group is such 
an active trypanocidal radicle, and we are accustomed to call it the 
‘ trypanophobe * group. 

For our starting point we took ‘ Phenolphthaleine,’ V, which on 
being injected into the alkaline tissues, changes into, VI, a Triphenyl- 
methane. This compound is of the same character as Malachite- 
Green and contains no NH 2 groups; we found it to be without effect 
upon the trypanosomes. We then proceeded to ‘ Fluorescein,’ VII, 
and to different halogenic fluorescein derivatives, such as Eosin, 
Floxin, &c.; here, too, there was no trypanocidal effect. But when 
‘ Rhodamine/ VIII, was used, it was found to have a very distinct 
effect upon the parasites; it possesses NH 2 groups. 



2J2 



V. THE THERAPEUTIC VALUES OF ATOXYL AND ACETYLATED 
ATOXYL COMPARED (8) 

Under this heading experiments were made with dogs, guinea 
pigs, and mice, and the effects of Atoxyl, of acetylated Atoxyl, and 
of acetylated Atoxyl followed by Mercury bichloride were compared. 
Trypanosoma brucei was the infecting parasite in each instance. 

A. Dogs. 

i. Treatment by Atoxyl alone. 

Experiment 415.—Trypanosomes appeared in the peripheral circulation on the 
fourth day after inoculation ; 5 c.cm. of a two per cent, solution of Atoxyl was 
given on the fifth day. The trypanosomes disappeared from the blood, but 
death ensued on the seventh day. 

Experiment 421.—Trypanosomes appeared in the peripheral circulation on the 
fourth day after inoculation. A dose of 2 c.cm. of a five per cent, solution of 
Atoxyl was given on the fourth day and again on the fifth day. Death followed 
on the eighth day although the parasites had disappeared and remained absent. 

Experiment 431.—Trypanosomes appeared in the peripheral blood on the 
fourth day. On the fourth and again on the fifth day a dose of 3 c.cm. of a five 
per cent, solution of Atoxyl was given. The parasites disappeared but reappeared 



on the twelfth day; a third dose of the same solution of Atoxyl was then given, 
but death followed on the fourteenth day of the experiment although-the parasites 
were not seen. 

Experiment 436.—Trypanosomes appeared in the blood on the fourth day. 
On the fifth and again on the sixth day a dose of 3 c.cm. of a five per cent, 
solution of Atoxyl was given. The trypanosomes disappeared and remained absent, 
but death followed on the tenth day of the experiment. 

Experiment 437.—Trypanosomes appeared in the peripheral circulation on the 
fourth day; a dose of 3 c.cm of a 5 per cent, solution of Atoxyl was given on the 
fifth day and on the sixth. The parasites disappeared and remained absent but 
the animal died on the ninth day. 

From these five experiments it may be concluded that dogs 
infected with the strain of Trypanosoma brucei employed would die, 
though treated by Atoxyl, in about nine days. 

2. Treatment by acetylated Atoxyl alone. 

Experiment 416.—Trypanosomes appeared in the peripheral circulation 
on the fourth day. On the fifth day 5 c.cm. of a two per cent, solution of 
acetylated Atoxyl was given. The trypanosomes did not disappear so this dose was 
repeated on the seventh day; it was again repeated on the twelfth day, 

although the parasites had disappeared after the second dose. They 

reappeared on the thirtieth day; two doses, each of 5 c.cm. of a three per cent, 
solution were therefore given on the thirtieth and thirty-first days. The parasites 
disappeared after the last of these doses and did not reappear until the forty-fourth 
day, when two doses, each of 5 c.cm. of a three per cent, solution, were given on 
successive days and the parasites disappeared, to reappear once more on the 
fifty-sixth day. In spite of another dose then given of 5 c.cm. of a three per 
cent, solution, the animal died on the following day. Rats subinoculated on the 
fifteenth and thirty-seventh days remained negative 

A rat subinoculated on the fifty-first day, five days before death, became 

injected , however, after a prolonged incubation period of eleven days and died 

four days later. 

Experiment 419.—Trypanosomes appeared in the peripheral circulation on the 
fourth day. On the fifth and again on the sixth day a dose of 5 c.cm. of a two per 
cent, solution of acetylated Atoxyl was given. The parasites disappeared but 
recurred on the eleventh day when two more doses, each of 5 c.cm. of the same 
solution were administered; although the parasites again disappeared, the dog 
died on the eighteenth day after inoculation. 

Two dogs treated by acetylated Atoxyl alone lived eighteen and 
fifty-seven days. 

3. Treatment by acetylated Atoxyl followed, by bichloride 

of Mercury . 

Experiment 334.—Trypanosomes appeared in the blood on the second day 
after inoculation. On the third, fourth and fifth days three doses of 5 c.cm. of a 
ten per cent, solution of acetylated Atoxyl were given; on the sixth and seventh 
days two doses of 10 c.cm. of the same solution were administered. On the 
eighth, the ninth and the tenth days 2 c.cm. of a one per cent, solution of Mercury 
bichloride were injected, and on the eleventh day were followed by a dose of 



2 74 


5 c.cm. of a ten per cent, solution of acetylated Atoxyl. The parasites disappeared 
from the blood on the fifth day of the experiment and were absent until the death 
of the animal on the forty-seventh day. 

A rat was inoculated on the seventh and another on the twenty-fourth day of 
the experiment; neither of them ever became infected. 

Experiment 420.—Trypanosomes appeared in the peripheral blood on the third 
day after inoculation. On the fourth and again on the fifth day 2 c.cm. of a 
five per cent, solution of acetylated Atoxyl was given. The parasites disappeared, 
and although they were still absent on the ninth and three succeeding days, 
13 c.cm. of the same solution was given in four doses of increasing size ; no toxic 
symptoms were observed ; and on the fourteenth, and again on the fifteenth day 
2 c.cm. of a 0*5 per cent, solution of Mercury bi-chloride was given. The 
parasites still remained absent, when on the twenty-ninth and thirtieth days two 
more doses of 5 c.cm. of the same solution of acetylated Atoxyl were injected and 
were followed on the two next days by two doses of 3 c.cm. of the same solution 
of Sublimate. The parasites remained absent until the death of the animal on the 
one hundred and fifth day; death was due in a large measure to very extensive 
mange. 

On the twentieth, forty-first, and sixty-seventh days of the experiment 
respectively, two rats and one puppy were inoculated with large quantities of 
blood ; none of them ever became infected. 

Experiment 432.—Trypanosomes appeared in the blood on the second day. 
On the third and again on the fourth day 3 c.cm. of a five per cent, solution of 
acetylated Atoxyl was given ; a dose of 5 c.cm. of a ten per cent, solution of 
acetylated Atoxyl was injected on the tenth and again on the eleventh day. This 
dose was repeated on the seventeenth and eighteenth days and was then followed 
on the nineteenth and twentieth days by two doses, each of 2 c.cm. of a 0-5 per 
cent, solution of Mercury bichloride. The parasites disappeared from the blood 
on the fourth day of the experiment and never reappeared. The animal died on 
the thirty-seventh day. 

A rat was subinoculated from this animal on the sixteenth day ; a rat and two 
mice on the twenty-eighth day, and a rat on the thirty-fifth day ; none of these 
animals ever became infected. 

Three dogs treated by acetylated Atoxyl lived from thirty-seven 
to one hundred and five days. 

B. Guinea Pigs. 


I. Treatment by Atoxyl alone. 

Experiment 406a.—Trypanosomes appeared in the blood on the fourth day. 
On the fifth day 1 c.cm. of a five per cent, solution of Atoxyl was given and the 
parasites disappeared, but death ensued on the seventh day. 

Experiment 406b.—Trypanosomes appeared in the blood on the sixth day. On 
the seventh and again on the eighth day 1 c.cm. of a five per cent, solution of 
Atoxyl was given. The parasites disappeared, but the animal died on the ninth 
day. 

Experiment 406c.—Trypanosomes appeared in the blood on the eighth day. 
On each of the two following days 1 c.cm. of a five per cent, solution of Atoxyl 
was given, and the parasites disappeared from the blood, but death followed on 
the fifteenth day. 

Three animals treated by Atoxyl alone died in from seven to 
fifteen days. 



2 75 


2. Treatment by acetylated Atoxyl followed by bichloride 

of Mercury. 

Experiment 407b.—Trypanosomes appeared in the blood on the sixth day. 
On the twelfth, thirteenth, twenty-second and twenty-third days, doses of 1*5 c.cm. 
of five per cent, solution of acetylated Atoxyl were given. These were followed on 
the twenty-fourth, twenty-fifth and twenty-sixth days by 0*5 c.cm. of a two per 
cent, solution of bichloride of Mercury. The animal died on the twenty-seventh 
day. No trypanosomes had been seen in the blood since the first dose of Atoxyl. 

Experiment 407c.—Trypanosomes appeared in the blood on the eighth day. 
On the twelfth and thirteenth days 2 c.cm. of a five per cent, solution of acetylated 
Atoxyl were given and were immediately followed on the two succeeding days by 
doses of 1 c.cm. of a two per cent, solution of Mercury bichloride. The parasites 
disappeared after the first dose of Atoxyl and did not reappear, but the animal 
died on the sixteenth day 

Experiment 414a.—Trypanosomes appeared in the blood on the eighth day. 
On the tenth, eleventh, and twelfth days, doses of 1*5 c.cm. of a five per cent, 
solution of acetylated Atoxyl were given. These were followed on the thirteenth, 
fourteenth, fifteenth, and sixteenth days by doses of 0*5 c.cm. of a two per cent, 
solution of Mercury bichloride, and on the seventeenth and eighteenth days by 
3*5 c.cm. of the same solution of acetylated Atoxyl in two doses, and, finally, on the 
nineteenth and twentieth days, by two doses of 1 c.cm. of the .same solution of 
Mercury bichloride. The trypanosomes disappeared from the blood on the second 
day of treatment and were still absent at the death of the animal from pneumonia 
on the sixty-seventh day. 

Rats were subinoculated on the fourteenth, fortieth, and fifty-fourth days of 
the experiment; none of them ever became infected. 

Experiment 414.—Trypanosomes appeared on the third day. Treatment was 
commenced on the eleventh day by five consecutive daily doses of 1-5 c.cm. of a 
five per cent, solution of acetylated Atoxyl. These were followed on the seventeenth 
and eighteenth days by two doses of 0*5 c.cm. of a two per cent, solution of 
bichloride of Mercury. The parasites disappeared from the blood on the second 
day of treatment and remained absent, but the animal died on the nineteenth day. 

Experiment 419.—Trypanosomes appeared in the blood on the eighth day. 
Treatment was commenced on the thirteenth day with three consecutive doses of 
1*5 c.cm. of a five per cent, solution of acetylated Atoxyl. It was continued on the 
twentieth and twenty-first days by two doses of 2 c.cm. of the same solution of 
Atoxyl, followed on the three next days by daily doses of 0*5 c.cm. of a two per 
cent, solution of Mercury bichloride, and on the twenty-sixth day by one more dose 
of 2 c.cm. of Atoxyl. The parasites which had disappeared from the blood on the 
second day of treatment remained absent until the forty-fifth day. On their 
reappearance their treatment was resumed by four consecutive daily doses of 
2 c.cm. of the same solution of Atoxyl; these were followed on the fiftieth day by 
1 c.cm. of a two per cent, solution of Mercury bichloride. The animal died on the 
next day, trypanosomes being absent from its blood. 

Five guinea pigs treated by acetylated Atoxyl and bichloride of 
Mercury died in from sixteen to sixty-seven days (average about 
thirty days). 



276 


C. Mice. 

Only one experiment was made, with the object of comparing the 
relative therapeutic values of Atoxyl and acetylated Atoxyl. 

Experiment 443.—Two lots of three mice each were inoculated with 
approximately equal amounts of infected blood from a common source. All became 
infected on the third and fourth days. Each animal on the day it was found to 
be infected was treated ; three mice received Atoxyl, and three acetylated Atoxyl 
(approximately equal quantities of the drug were given). Those treated with 
Atoxyl died on the fourth and fifth days; those with acetylated Atoxyl on the 
ninth and eleventh days. 

From these experiments, so far as they permit comparisons and 
conclusions, we deduce for the animals concerned, that: — 

Treatment by acetylated Atoxyl followed by Mercury is more 
efficacious than is treatment by acetylated Atoxyl alone; acetylated 
Atoxyl is of more value than Atoxyl; but that none of these methods 
is of practical value since death invariably occurred.* 


VI. TREATMENT BY ATOXYL FOLLOWED BY ANOTHER DRUG 

As explained in our former paper, it was thought that substances, 
ordinarily without therapeutic value, might be trypanocidal when 
administered after Atoxyl; it was this line of work which led to the 
discovery of the efficacy of Atoxyl combined with bichloride of 
Mercury in the treatment of trypanosome-infected rats. 

The following combinations were tried and found to be valueless ; 
the parasites generally reappeared in the blood in from three to four 
weeks after the cessation of treatment, and death followed in due 
course. 

Atoxyl and Silver Nitrate. 

Atoxyl and Lead Acetate. 

Atoxyl and Quinine-cacodylate. 

Atoxyl and Potassium Bichromate. 

Atoxyl and Quinine. 


* It is true that trypanosomes could not be detected, either by subinoculation 
or direct examination, in some of these animals for many days before their death, 
and it must be asked whether they really died of trypanosomiasis or from some other 
cause, intoxication from the treatment for example ; gross signs of overdosage were 
not observed. The same question must be asked concerning the deaths of many 
of the animals experimented with; this is particularly the case with the donkeys. 
Experiments 416 and 419 show definitely how infection may declare itself after 
being long dormant in thoroughly treated animals. 



277 


VII. TREATMENT OF ATOXYL FOLLOWED BY BICHLORIDE OF 

MERCURY ’ 

Under this heading are reported our attempts to apply the 
combined use of Atoxyl and bichloride of Mercury, found successful 
in the treatment of rats, to the cure of large animals infected with 
trypanosomes. Rabbits and donkeys were used in this series of 
experiments. 

A. Rabbits. 

i. Controls , treated by Atoxyl alone. 

Three control rabbits which only received the same amount of 
Atoxyl as was used in the combined treatment by Atoxyl and 
bichloride of Mercury, all died in about fifty-five days after inocula¬ 
tion. In one rabbit (Experiment 302) death also occurred, although 
the dosage by Atoxyl was repeated continuously. In all, the 
infecting trypanosome was Trypanosoma brucei. 

Experiment 302.—Fourteen days after inoculation trypanosomes appeared in 
the blood. Treatment was commenced nine days later. On the first day 1 c.cm. of a 
five per cent, solution of Atoxyl was given; one week later another dose of 1 c.cm. 
was injected, and on the day following 2 c.cm. The dosage of 2 c.cm. was then 
repeated for ten times at intervals of from five to eight days; so that in all 23 c.cm. 
of Atoxyl solution were given in twelve doses in forty-two days to a rabbit weighing 
1805 grammes at the commencement of the experiment. Trypanosomes reappeared 
in the peripheral blood iii forty-three days after the commencement of treatment, 
and the animal died three days later weighing 1560 grammes. 

Rabbits treated by Atoxyl alone, whether continuously or not, 
died in about fifty to sixty days after inoculation. 

2. Treatment by Atoxyl followed by bichloride of Mercury. 

(a) Rabbits infected with Trypanosoma brucei 

Experiment 288.—After fifteen days trypanosomes appeared in the blood of a 
rabbit inoculated on January 7. Six days later treatment was commenced, the 
animal had already lost no grammes of an original weight of 2350 grammes. 
Three c.cm. of a five per cent. Atoxyl solution were given in four doses during a 
period of eleven days; 2 c.cm. being given in the last three days. Three day9 after 
the last dose of Atoxyl the dosage by bichloride of Mercury was commenced, and 
4 c.cm. in four doses were given in five days’ time. The blood was examined daily 
for one hundred and forty-seven days, until January 10th, when the animal died. 
No macroscopic cause of death was seen at the autopsy; there were no signs of 
trypanosomiasis. Mice were subinoculated from this animal one day after treatment 
was stopped and at fourteen, twenty-five, fifty-one and one hundred and forty-one 
(3 mice) days from that date. A young puppy was inoculated (with 10 c.cm. of 
blood) at the time of the autopsy. All these animals were carefully observed for a 
sufficient length of time; none of them were ever observed to become infected with 
trypanosomes. A rat inoculated on the third day after the first dose of Atoxyl 
became infected in nine days and died three days later. 



278 


Experiment 289.—Six days after its inoculation on January 24th, trypanosomes 
appeared in the peripheral blood of an adult female rabbit. Treatment was com¬ 
menced five days later, and 3*5 c.cm. of five per cent. Atoxyl were given in four 
doses on four successive days. On the four following days 4 c.cm. of a two per cent, 
solution of Mercury bichloride was given in four equal doses. In spite of very 
careful examination trypanosomes could never be found in the peripheral blood of 
the animal, although it steadily lost weight; thus at the time of inoculation it 
weighed 2020 grammes, at the commencement of treatment 1850 grammes, and at 
seventy days after the cessation of treatment 1712 grammes. A puppy was sub¬ 
inoculated at this time but it never became infected. Because of this loss 
of weight a second course of treatment was commenced eighty-two days after 
the stoppage of the first course; 8 c.cm. of Atoxyl was given in three approximately 
equal doses on three successive days and was immediately followed on the two 
next days by two doses, each of 2 c.cm., of a two per cent, solution of bichloride 
of Mercury. The animal’s weight soon commenced to increase, and in six weeks 
160 grammes was gained. The animal, unfortunately, died from pneumonia on 
August 8th, two hundred and seventy days after inoculation. Its blood had been 
examined almost daily and trypanosomes were never seen, nor were they ever found 
in a rabbit and two mice subinoculated with large amounts of blood from it on 
July 12th and July 28th respectively. 

Experiment 291.—An adult male rabbit was inoculated on January 24th, 
1Q07. After ten days trypanosomes appeared in its blood. Four days later treat¬ 
ment was commenced and 4 c.cm. of five per cent. Atoxyl solution was given in equal 
doses during five days. This was immediately followed on four successive days 
by 4c.cm., given in four equal doses, of a two per cent, solution of bichloride of 
Mercury. The animal’s blood was examined carefully, but trypanosomes were 
never again seen, and the animal was still living on the 3rd March, 1908, four 
hundred and three days after inoculation. 

Rats were subinoculated from this animal at fourteen, forty-five, seventy-five, 
one hundred and six, one hundred and thirty-six, and one hundred and sixty-seven 
days after the cessation of treatment. All of them have been carefully examined 
and none of them have ever been found to be infected. 

Experiment 350.—Trypanosomes were first seen in the blood of an adult 
female rabbit on April 29th, 1907, twenty-five days after its inoculation. Treatment 
was commenced next day, on April 30th, and 10 c.cm. of Atoxyl solution was given 
in four doses during the next seven days; 6 c.cm. were given in the last two doses. 
This was immediately followed by 4 c.cm. of a two per cent, solution of bichloride 
of Mercury, given in four equal doses on four successive days. The animal was 
still alive on March 3rd, 1908, three hundred and thirty-three days after 
inoculation. Trypanosomes were never seen in its blood, nor did they ever appear 
in the blood of rats subinoculated from it on the last day of treatment, and at 
fifty-eight, eighty-nine, and one hundred and twenty days from that date. 

Experiment 356.—Trypanosomes appeared in the peripheral blood of an adult 
female rabbit seven days after its inoculation. Six days later treatment w r as 
commenced, and 2 c.cm. of a ten per cent, solution of Atoxyl was given in one 
dose. This dose was repeated seventeen days later, and then was immediately 
followed by three equal doses of 1 c.cm of a two per cent, solution of Mercury 
bichloride, given on three successive days. Trypanosomes were not seen in the 
animal’s blood from the date of the first injection of Atoxyl to its death, forty-six 
days later (no autopsy; cause of death unknown), nor were trypanosomes seen in 
the blood of a rat subinoculated from it at thirty-one days from the commence¬ 
ment of treatment. 

Of five rabbits infected with Trypanosoma brucei and treated by 



279 


Atoxyl followed by bichloride of Mercury, one died in forty-six days 
and four were apparently cured ; this combined treatment of rabbits 
infected with Trypanosoma brucei therefore is much superior to the 
treatment by Atoxyl alone, and will sometimes effect a cure. 

( b ) Rabbits infected with Trypanosoma 
gambiense 

Experiment 323.—Trypanosomes appeared after forty-one days in the blood 
of a rabbit inoculated on February 26th. Treatment was commenced sixteen days 
later and 7 c.cm. of five per cent. Atoxyl solution was given, in four doses at equal 
intervals, during the next twelve days. This was immediately followed during 
the succeeding ten days by 4 c.cm. of bichloride solution, given in four equal 
doses at approximately equal intervals. The rabbit's blood was examined 
practically daily for one hundred and forty-one days, until October 5th, when it 
died (cause not known ; no autopsy). Trypanosomes were never seen in it after 
the treatment was stopped, nor in mice subinoculated from it thirty days from 
that time 

Experiment 326.—After eighteen days trypanosomes were found in the ear 
blood of a rabbit inoculated on February 26th, 1907. The parasites were con¬ 
stantly present when treatment was commenced forty-three days later, and 8 c.cm. 
of five per cent. Atoxyl solution was given in four doses at approximately equal 
intervals during the next fifteen days. This was immediately followed by 4 c.cm. 
of Mercury bichloride solution, given in four doses at almost equal intervals 
during the succeeding ten days. The rabbit’s blood was examined daily for 
forty-nine days and at intervals of two or three days until January 26th, 1908, 
eleven months after inoculation, when it was Killed. Trypanosomes were never 
seen in it after treatment was stopped, nor in the three rats and two mice sub¬ 
inoculated from it just at the end of treatment and a month after treatment had 
stopped. 

Experiment 327.—After thirty-one days trypanosomes were found in the 
peripheral blood of an adult male rabbit inoculated on February 22nd, 1907. Treat¬ 
ment was commenced thirty days later, and 8 c.cm. of five per cent. Atoxyl solution 
was given in four doses during the next twelve days. This was followed in the 
succeeding ten days by 4 c.cm. of one per cent. Mercury bichloride solution, 
given in four equal doses at almost equal intervals of time. The rabbit’s blood 
was examined daily after the cessation of treatment for one hundred and eighty- 
four days and, after that, at intervals until February 3rd, 1908, over eleven months 
after inoculation, when the animal was killed because of severe skin disease. 
Trypanosomes were never seen in its blood after the treatment was stopped, nor 
in mice and rats subinoculated, respectively, a month after the stoppage of 
treatment and at the autopsy. 

Experiment 329.—After twenty-six. days trypanosomes appeared in the 
peripheral blood of a rabbit. Treatment was commenced thirty-two days later, 
and 11 c.cm. of five per cent. Atoxyl solution was given in five doses during the 
next twelve days; just over half of the drug was given in the last two days. This 
was followed during the next nine days by 7 c.cm. of bichloride solution, given in 
four doses; just half of this drug was given on the first two days. After cessation 
of the treatment, the rabbit’s blood was examined daily for one hundred days, when 
it died (cause not known ; no autopsy). Trypanosomes were never seen in its 
blood, nor did they appear in the blood of a mouse inoculated at the termination 
of the treatment nor in rats inoculated at respectively thirteen and seventy-four 
days from that time. 



28o 


The disease produced in rabbits by Trypanosoma gambiense may 
run a very chronic course; it is therefore better to say nothing 
concerning Experiments 323 and 329. Experiments 326 and 327 
may, however, be safely considered to have been cured of an infection 
by Trypanosoma gambiense. 

B. Donkeys. 

The outcome of this series of experiments was extremely 
disappointing. It was commenced immediately after the favourable 
results had been obtained in the treatment of rats by means of 
combined Atoxyl and Mercury bichloride. There is no doubt that in 
the first of the succeeding experiments the disease was allowed to 
go too far before the commencement of treatment; but even when, 
as in the later experiments, treatment was commenced early, it was 
found impossible to save a single animal. 

In all the donkey experiments, as a general rule, the temperature, 
taken twice daily, became higher when the trypanosomes reappeared 
in the peripheral blood; whether trypanosomes were present or not 
it was often noticed that an elevated temperature fell immediately 
after the administration of another dose of the drug, and it was 
frequently observed that animals inoculated at such a time, that is, 
when the examination of the blood was negative and the temperature 
was high, often became infected. As a rule, therefore, a high 
temperature was accepted as a definite indication for a dose of Atoxyl. 

1. Controls , treated by Atoxyl alone. 

The disease was allowed to run its course, without treatment, in 
two donkeys; trypanosomes appeared in the blood on the third day, 
and they died after a typical illness, with remitting and intermitting 
fever going up to 104° or 105° F., in eighteen and twenty-four days 
respectively; at the post-mortem typical signs of trypanosomiasis 
were present. 

Treatment was commenced on the twentieth day in a third 
animal, inoculated at the same time, when it was already practically 
moribund ; it died on the twenty-third day. 

Experiment 9.—Trypanosomes appeared four days after inoculation. They 
increased in numbers slowly and treatment was not commenced until the twenty- 
sixth day, when 10 c.cm of a two per cent, solution of Atoxyl was given. This 
dose was repeated on the thirty-second and thirty-third days because of a recurrence 



of the trypanosomes, which had disappeared after the first inoculation. The 
parasites then again left the blood, only to reappear on the forty-first day. Death 
followed on the forty-sixth day. 

Experiment 391.—Trypanosomes appeared in the blood on the fifth day after 
inoculation. On the thirty-first day treatment was commenced by a dose of 
ioc.cm. of a ten per cent, solution of Atoxyl. This was repeated, as the parasites 
reappeared in the blood, on the thirty-second, thirty-third, thirty-eighth, forty- 
second, and forty-fifth days. Since the last dose of Atoxyl did not drive out the 
trypanosomes, a dose of 20 c.cm. of a twenty per cent, solution of Atoxyl was 
given. The parasites disappeared then from the blood, but the animal died three 
days later on the forty-eighth day. 

Experiment 393.—Trypanosomes appeared in the blood on the fourth day. 
Treatment was commenced on the thirty-fifth day by giving four consecutive daily 
doses of 10 c.cm. of a ten per cent, solution of Atoxyl. The parasites disappeared 
from the blood, only to reappear on the forty-sixth day, when 10 c.cm. of a twenty 
per cent, solution of Atoxyl was given. The parasites once more disappeared to 
return again on the fifty-eighth day, when a dose of 20 c.cm. of a twenty per cent, 
solution of Atoxyl was given. The parasites again disappeared from the blood 
and did not return before death on the sixty-seventh day. Three rats were sub¬ 
inoculated from this animal on the thirty-seventh, sixty-first and sixty-sixth days, 
when trypanosomes were not present. All three of these animals were carefully 
examined for periods of from three to four months ; none of them ever became 
infected. 

Experiment 2. —Trypanosomes appeared in the blood on the fourth day. 
Treatment was commenced on the thirty-fourth day by 10 c.cm. of a ten per cent, 
solution of Atoxyl. This dose was repeated daily on the three following days and 
again, when twice that amount was given, on the forty-fourth day. But the 
trypanosomes, which had been absent since the second dose, reappeared on the 
forty-seventh day and the animal, in spite of a dose of 10 c.cm. of a ten per cent, 
solution of ‘Afridol,’ died on the day following. Rats subinoculated on the 
thirty-sixth and forty-sixth days never became infected. 

Experiment 393.—Trypanosomes appeared in the blood on the fourth day. 
Treatment was commenced on the twenty-sixth and two following days by giving 
daily 10 c.cm. of a ten per cent, solution of Atoxyl. In spite of the most careful 
nursing, the animal died on the thirty-seventh day, although the trypanosomes did 
not reappear in the peripheral blood. Two rats subinoculated on the thirty-first 
day of the experiment never became infected. 

Experiment 17.—Trypanosomes appeared in the blood on the third day. 
Treatment consisted of 10 c.cm. of a ten per cent, solution of Atoxyl, given on 
the twenty-second, twenty-seventh and twenty-eighth days and, thereafter, once a 
week until the death of the animal on the ninety-seventh day. The trypanosomes 
disappeared from the blood immediately after the first dose of Atoxyl, and did not 
reappear until the seventy-eighth and seventy-ninth days, when they again dis¬ 
appeared and were absent at death. The chart of this animal shows a typical high 
fever until the commencement of treatment; then the temperature remained 
approximately normal with slight rises until the seventy-sixth day, when it again 
became high and continued to run an elevated course. 

Experiment 19.—Trypanosomes appeared in the peripheral blood on the third 
day. Treatment was commenced on the twenty-fourth dav. when 10 c.cm. of a 
ten per cent, solution of Atoxyl was given. This dose was repeated on every 
second day from that time until the death of the animal on the seventy-third day. 
Trypanosomes were not seen in the blood after the first dose of Atoxyl. Rats 
inoculated on the twenty-fifth and thirty-eighth days of the experiment, however, 
became infected after a prolonged incubation of about two weeks and died in due 

U 



282 


course. The temperature of this donkey, although irregularly febrile, did not run 
as high as was usually the case ; it did not go above 103 0 after treatment was 
commenced. 

Experiment 14.—Trypanosomes appeared in the blood on the fifth day. 
Treatment was commenced on the seventeenth day by injection of 10 c.cm. of a 
ten per cent, solution of Atoxyl. ThTs same dose was repeated daily—Sundays 
excepted—from then until the ninety-fourth day of the experiment. The trypano¬ 
somes disappeared from the blood after the first dose of Atoxyl and did not 
reappear until the seventy-fourth and seventy-fifth days, when they once more 
disappeared until the ninety-fifth day. They disappeared once more and were not 
seen again before death on the one hundred and fourth day. Rats subinoculated 
just before the second dose of Atoxyl never became infected, while rats inoculated 
on the fifty-fourth and sixty-eighth days both became infected after incubation 
periods slightly longer than usual and died in due course. 

From these experiments it appears that daily doses of 10 c.cm. 
of a ten per cent, solution of Atoxyl, and occasionally twice that 
amount, could be given to an adult donkey without exciting acute 
signs of poisoning; but even these large doses were insufficient to 
keep the trypanosomes out of the blood and the animals invariably 
died, whether the parasites were to be found in the blood or no, in 
from twenty-three to one hundred days, usually in about fifty days. 

2. Treatment by Atoxyl and bichloride of Mercury. 

(a) Symptomatic treatment 

In the following series of experiments treatment was given 
practically only when it seemed necessary because of the presence of 
trypanosomes, or of a high temperature. 

At first .weaker solutions of Atoxyl were used; as it became 
apparent that they were inefficacious they were strengthened, until 
the saturated twenty per cent, solution was currently employed ; 
this same strength of the solution was used in all the later 
experiments. 

Experiment it.— Trypanosomes appeared in the blood five days after 
inoculation. On the eighth and ninth days treatment was commenced by the daily 
injection of 10 c.cm. of a five per cent, solution of Atoxyl; on the twelfth and 
thirteenth days, doses of 1 c.cm. of a one per cent, solution of Sublimate were 
given; on the fourteenth day four times that amount of the drug was administered. 
On the fifteenth, eighteenth, twenty-ninth, thirty-fourth and thirty-fifth days 
10 c.cm. of Atoxyl was injected. The trypanosomes disappeared from the blood 
on the second day of treatment; there were one or two considerable elevations 
in temperature but the parasites remained absent from that time until the twenty- 
ninth day, when 10 c.cm. of a ten per cent, solution of Atoxyl was given for three 
days, and was immediately followed by 10 c.cm. of a two per cent, solution of 
bichloride of Mercury. The parasites disappeared from the blood as a result of 
this treatment, but reappeared on the forty-sixth day. Three daily doses of 10 c.cm. 



283 

of a ten per cent, solution orf Atoxyl were then given and the parasites once more 
disappeared, only to reappear later and remain more or less constantly present 
until the death of the animal on the eighty-third day. 

Experiments 8, to and 12.—Only the first of these experiments is given in 
full. The treatment was practically identical in all. The course of the disease 
was much the same and the same disappointing result ensued in each. 

Experiment 8. —See Chart I. 

Experiment 10.—Trypanosomes appeared in the blood on the fourth day. 
Treatment was commenced on the eleventh day. The parasites reappeared in 
the blood after a prolonged absence on the thirty-first day and again at intervals 
until the death of the animal at one hundred and seventy-five days. 

It is interesting to note that a rat subinoculated two days before the 
recurrence of the trypanosomes in the early part of the infection remained 
negative, while another rat inoculated during a very slight rise in temperature 
(102 0 ) at a time when no trypanosomes were present in the blood for ten days 
before or after, became infected. 

Experiment 12.—Parasites appeared in the blood on the fourth day. Treat¬ 
ment was commenced on the eleventh day. The parasites were present in the 
blood at long intervals until the death of the animal at two hundred and three days. 

Subinoculations were made from this animal on the twenty-ninth, forty- 
*fifth, ninety-first, and one hundred and first days. That made on the forty-fifth 
day was negative, although trypanosomes had been present in the blood only four 
days previously; those made on the twenty-ninth and one hundred and first days 
were also negative, while that made on the ninety-first day, although trypanosomes 
were not seen in the blood for approximately a month before or after, was 
positive. The temperature was not elevated at this time. 

Experiment 18.—Trypanosomes appeared in the blood after three days. 
Treatment was commenced on the eighth day by the injection of 10 c.cm. of a 
twenty per cent, solution of Atoxyl. This dose was repeated on the following day 
and on the seventeenth and eighteenth days, while one dose of 5 c.cm. of a five 
per cent, solution of bichloride of Mercury was given on the tenth day. The 
trypanosomes only left the blood for two days after the first dose of Atoxyl, they 
then reappeared. In spite of the repeated doses of Atoxyl they persisted, and the 
animal died on the nineteenth day. 

The early persistence of the parasites, after vigorous treatment, 
in this experiment is extraordinary; they seem to have suddenly 
acquired Atoxyl-resistant characters. Such an occurrence is unique 
in our experience. 

Experiments 22-26.—In this series of experiments the dose of Atoxyl was 
always 10 c.cm. of a twenty per cent, solution, and the dose of bichloride 10 c.cm. 
of a two per cent, solution. As a rule a series of two doses of Atoxyl followed by 
one of bichloride of Mercury was given weekly on three successive days. If 
trypanosomes appeared in the blood or if the temperature were high, this dose 
was repeated at closer intervals, while if the temperature were normal and the 
parasites absent it was occasionally omitted for one week. 

Experiment 25.—This experiment is given in full (Chart II.). It is a typical 
example of the course and fatal issue of the disease in the remaining experiments. 

Experiment 22.—The parasites appeared in the blood in two days. Treat¬ 
ment was commenced at once. The temperature ran an irregular course and the 
parasites appeared at irregular intervals, until death ensued on the seventy-ninth 
day. 



284 


Experiment 23.—Trypanosomes appeared in the blood on the second day. 
Treatment was commenced on the seventeenth day. The parasites, which 
recurred early in the treatment, were absent for six weeks toward its end, but they 
eventually reappeared and the animal died on the one hundred and twenty-second 
day. 

A rat subinoculated on the twenty-eighth day became infected after an 
inoculation period of 23 days, although trypanosomes had not been seen in the 
blood for a week and the treatment of three doses of Atoxyl and two of Mercury 
bichloride had only ended five days previously. The temperature was not high 
(99*2°), and trypanosomes did not appear in the blood for two weeks later. 

Experiment 24.—Parasites appeared in the blood on the third day. Treat¬ 
ment was commenced four days later. The parasites were very persistent at the 
first part of this experiment, although they were absent during the last three weeks. 
The animal died on the thirty-third day. 

Two rats subinoculated while the parasites were absent, on the day of death 
and ten days previously, both became infected. 

Experiment 26.—Parasites appeared on the twenty-second day. Treatment 
commenced three days later. Death followed on the forty-second day. 

( b ) Routine treatment 

Since symptomatic treatment Tailed, the continuous, routine 
administration of large doses of both drugs was adopted as a last 
resort; it, too, failed. 

Experiments ib and 21.—The treatment after the commencement was the same 
in both these animals. A dose of 10 c.cm. of a twenty per cent, solution of Atoxyl 
was given on the first and second days, and was followed by 10 c.cm. of a two per 
cent, solution of bichloride of Mercury on the third day. This treatment was 
repeated twice weekly; on Sundays no treatment was given. 

Experiment 16.—Trypanosomes appeared in the blood on the fourth day. 
Treatment was commenced on the twenty-fourth by three successive doses of 
10 c.cm. of a ten per cent, solution of Atoxyl. Treatment as described above was 
thenceforth continued until the one hundred and sixth day, when the parasites, 
which had been continually absent since the second day of treatment, reappeared. 
Trypanroth in daily doses of 10 c.cm. of a one per cent, solution was then given 
on six consecutive days. The parasites failed to disappear and the animal died 
on the one hundred and twentieth day. 

Animals subinoculated on the fiftieth day did not become infected, while one 
inoculated on the ninety-third day, thirteen days before the reappearance of the 
parasites, became infected after an incubation of seven days. 

Experiment 21.—Trypanosomes appeared in the blood on the third day. Treat¬ 
ment was commenced on the twenty-sixth day, and in the next fourteen days 
seven doses of 10 c.cm. of a ten per cent, solution of Atoxyl were given. The 
parasites, which disappeared after the first two doses of Atoxyl, reappeared twice 
during this course of meejication. The Atoxyl was followed by 16 c.cm. of a two 
per cent, solution of Sublimate, given in three doses, on two consecutive days. 
From the forty-sixth to the one hundred and fifth day the treatment indicated 
above was given regularly. The parasites disappeared at once and remained absent 
to examination and subinoculation until the ninetieth day, when they once more 
reappeared and were constantly present until death on the one hundred and 
twentieth da}*. 

It is interesting to note that a rat subinoculated with blood taken during the 
first course of treatment with Atoxyl, although trypanosomes were present, only 
became infected after an incubation of sixteen days. 



2**5 


Experiment 20. —Trypanosomes appeared on the fourth day. O11 the sixth, 
treatment was commenced and was continued until two days before death on the 
thirty-eighth day. For two days 10 c.cm. of a twenty per cent, solution of Atoxyl 
was given daily and on the third day 10 c.cm. of a two per cent, solution of 
bichloride of Mercury was administered. This treatment was repeated twice 
weekly. The trypanosomes disappeared from the blood five days after treatment 
was commenced and never reappeared. 

A rat subinoculated on the thirty-fourth day, four days before death, became 
infected after a long incubation of twenty-three days. Another rat subinoculated 
on the seventeenth day never became infected. 

Donkeys treated by Atoxyl followed by bichloride of Mercury all 
died in from about thirty-three to two hundred days (average one 
hundred days), in spite of large doses. 


VIII. TREATMENT BY TRYPANROTH FOLLOWED BY 
BICHLORIDE OF MERCURY 

This series of experiments was undertaken to ascertain whether 
Sublimate would have the same value when given after another 
trypanocidal substance as it possesses when following Atoxyl. There 
was a slight difficulty in carrying out these experiments, since 
Trypanroth was not always able to prevent an early death from the 
disease. 

Experiment 800.—Trypanosomes appeared in the blood of six rats on the first 
day after inoculation. A dose of 1 c.cm. of a one per cent, solution of Trypanroth 
was at once given and was repeated daily on the ten following days; the 
parasites had disappeared from each of the rats on the sixth day of the experiment. 
Three of the rats then received on the three following days doses of 0*5 c.cm. of 
a one per cent, solution of bichloride of Mercury; three received no further treat¬ 
ment. The parasites reappeared in all six rats and death followed in from two 
to four days. Those which received only Trypanroth died in sixteen, seventeen, 
and eighteen days; those which received Trypanroth and Sublimate in seventeen, 
nineteen, and twenty-two days. 

Experiment 802.—Trypanosomes appeared in six rats on the third day after 
inoculation. One c.cm. of a one per cent, solution of Trypanroth was at once given 
and was repeated on the six following days. The parasites disappeared from the 
blood on the eighth day of the experiment in five of the rats ; one died. Three 
doses, on consecutive days, of 1*5 c.cm. of a one per cent, solution of Mercury 
bichloride were then given to three of the rats; the remaining two had no further 
treatment. The parasites ultimately reappeared in all of them and death followed 
as usual in from two to four days. Those which received only Trypanroth died 
in fifteen and sixteen days from the commencement of the experiment; those which 
received the combined treatment in fourteen, twenty-five, and forty-eight days. 

From these experiments it seems that combined treatment by 
Trypanroth, followed by bichloride of Mercury, is superior to treat¬ 
ment by Trypanroth alone, but it is far inferior to the treatment by 
Atoxyl and Mercury. 



*86 


IX. SUMMARY AND CONCLUSIONS 

I. Certain definite rules which must be followed in the 
experimental therapeutics of trypanosomiasis are insisted upon. 

II. The after history of the rats dealt with in our first papers 
(*’ 2 ) is referred to. The Atoxyl-treated controls all eventually died 
of trypanosomiasis; those treated by Atoxyl followed by bichloride 
of Mercury, and by Atoxyl followed by Donovan’s solution, which 
were thought to be cured, have never had recurrences, but they were 
not immune to re-inoculation. 

Ill and VI. Several drugs and combinations of drugs found to be 
inefficacious in the treatment of infected rats are named. 

IV. None of the colouring matters employed were of much 
value ; it is suggested that the active radicle in trypanocidal aniline 
derivatives is the ‘ trypanophobe ’ group, NH a . 

V. In the treatment of dogs, guinea pigs, and mice, the 
comparative value of (i) acetylated Atoxyl followed by bichloride 
of Mercury, (2) of acetylated Atoxyl, and (3) of Atoxyl, is as the order 
in which they are named ; none of these methods is usually able to 
definitely cure well-established infections in these animals. 

VII. A. Atoxyl followed by bichloride of Mercury is found to 
be much superior to Atoxyl alone in the treatment of rabbits infected 
with Trypanosoma brucei ; the latter is also effective in the treatment 
of rabbits infected with Trypanosoma gambiense. 

B. Atoxyl and Mercury combined are distinctly superior to 
Atoxyl alone in the treatment of donkeys infected with Trypanosoma 
brucei , but neither method is able to cure a well-established infection. 

VIII. Treatment of rats infected with Trypanosoma brucei by 
Trypanroth followed by bichloride of Mercury is superior to treatment 
by Trypanroth alone, but inferior to the combined treatment by 
Atoxyl and bichloride of Mercury. 

IX. Many subinoculations were made during the work to ascertain 
whether the animals experimented with w^ere infected. The great 
majority of subinoculations made from animals in whose blood 
trypanosomes could not be seen were negative; thirteen, made under 
apparently identical circumstances, were positive. An examination 
of these subinoculations is interesting. 



287 


Rats became infected although trypanosomes were not seen in 
the animal from which the blood was taken for over a month both 
before and after the subinoculation. They became infected even 
although large doses of Atoxyl or of Atoxyl and Mercury had been 
given to the infecting animal only a day previously. Blood taken 
^during a rise in temperature was often infective; but blood taken 
while the temperature was no higher than usual, and at a time neither 
preceding nor following a rise, was also infective; blood taken 
during the customary ante-mortem fall of temperature was infective 
in four of these thirteen experiments, although trypanosomes were 
absent from it. 

The chief interest in these successful subinoculations lies in the 
fact that the incubation period was greatly lengthened; while the 
course of the disease, once the parasites appeared in the blood, was 
normal, and the rats died about three days later. 11 In six of these 
thirteen subinoculations the incubation period was over fifteen days, 
in three over twenty, the longest was twenty-six days; only once, 
and then in blood taken from a dying animal, was the incubation so 
short as four days. Although other explanations suggest themselves, 
these observations seem to be in harmony with our belief that 
recurrences in apparently-cured, trypanosome-infected animals are 
due to the production and persistence of some resistant developmental 
form of the parasite, and not merely to the acquirement of ‘ chemo¬ 
therapeutic resistant * properties by it. 



288 


REFERENCES 


1. Moore, Nierenstein and Todd. Concerning the treatment of experimental 

trypanosomiasis. Annals of Tropical Medicine and Parasitology, Vol. i. 
No. 2, June, 1907. 

2. Moore, Nierenstein and Todd. On the treatment of trypanosomiasis. 

13 io-Chemical Journal, Vol. II, Nos. 5 and 6, April, 1907. 

3. Ehrlich. Chemotherapeutische Trypanosomen-Studien. Rerliner klinische 

Wochenschrift, Nos. g-12,1907. 

4. Ehrlich and Shiga. Rerliner klinische Wochenschrift, Nos. 13 and 14, 1904. 

5. Mesnil and Nicolle. Traitement des trypanosomiases par les couleurs de 

benzidine. Annals de l’lnstitut Pasteur, T. XX, 1906. 

6. Wendelstadt and Eellmer. Ueber die Wirkung von Malachitgriin und 

anderen verschiedenartigen Stoffen gegen Nagana-Trypanosomen. Deutsche 
medizinische Wochenschrift, No 47, p. 1711, 1904. 

7. Nicolle and Mesnil. Traitement des trypanosomiases par les couleurs de 

benzidine. Annales de l’lnstitut Pasteur, T. XX, 1906. 

8. Ehrlich und Rertheim. Rerichte d. deutschen chem. Gesellschaft. Vol. 

40, pag. 3296. • 

9. Boyce and Breinl, and Manson. Annals of Tropical Medicine and 

Parasitology, Vol. II, No. 1, 1907. The literature of the whole subject of 
the treatment of trypanosomiasis by Atoxyl is reviewed in this number. 
Important papers of more recent date are (10) and (11). 

m. Laveran and Thiroux. Recherches sur le traitement des trypanosomiases. 
Annales de l’lnstitut Pasteur, T. XXII, f£vrier 1908. 

11. Browning. Chemotherapy in trypanosome infections. Journal of Pathology 

and Bacteriology, Vol. XII, 1908. 

12. Thomas and Breinl. Trypanosomes, Trypanosomiasis, and Sleeping Sick¬ 

ness: Pathology and Treatment. Memoir XVI of the Liverpool School of 
Tropical Medicine. 




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OOWKEY 






289 

AN UNUSUAL CASE OF GOUNDOU 


BY 

Dr. R. W. ORPEN, 

MEDICAL OFFICER, W.A.M.S., SIERRA LEONE 

(Received for publication 17 June , 1908J 

Whilst at Kanre Lahun, on the Liberian frontier, I chanced to 
see a case of Goundou which presented some unusual and interesting 
characters. 

The patient was a Mendi man, about 22 years of age. He stated 
that about five years ago the usual tumours began to grow, their 
appearance being preceded by a severe attack of yaws and constant 
severe headache. He said he had never had syphilis. About a year 
later, a third tumour began to develop on the left side of the face in 
the malar region. This third tumour is slowly increasing in size, 
whilst the others (nasal) have not increased to any appreciable extent 
during the past two years. It is only within the last few months that 
a discharge has been noticed from the right nostril, and no discharge 
has ever been observed from the left. Constant severe headache has 
been present, preventing him from working and causing loss of sleep. 
When he stoops down the pain is greatly increased. 

On examination the patient is seen to be of poor physique. 

Face. On each side of the nose, springing from the nasal 
processes of the superior maxillae, a tumour is seen. They are 
symmetrical, sessile, and of bony hardness. Somewhat oval in shape, 
the long axis being downwards and outwards, they are smooth with 
normal skin freely moveable over them. They are dull on 
percussion. There is a foul discharge coming from the right 
nostril. The passage of tears through the nasal ducts is not 
interfered with. The tumours practically fill the nostrils, and the 
patient is unable to breathe through the nose. The sense of smell is 
totally lost; he is unable to distinguish the odours of tobacco, 
ammonia, and peppermint. 

On the left side of the face a third tumour is seen in the malar 
region close to, but separate from, the previously described ones. In 
size and shape it is about that of half a hen's egg, but it is not 
entirely smooth and regular, as on its upper and outer aspect a 
protuberance can be felt. The long axis of the tumour is upwards 
and outwards, and the outer canthus of the eye is dragged with it. It 
w 



jgo 


does not invade the orbital cavity. The tumour is of bony hardness, 
is not painful on being handled, is dull on percussion, and the skin 
over it is normal and freely moveable. Pain is frequently felt 
radiating to the tempero-maxillary articulation. There is no sign of 
oedema or ‘ egg-shell * crackling. This tumour has developed more 
rapidly than the usual tumours. 

Examination of the mouth shows nothing abnormal, the patient 
possessing a perfect set of teeth with none missing. His palate is 
normal, though somewhat low, and both sides are symmetrical. His 
chest is flat; his breasts are unduly developed, and the nipples are 
large and prominent. His heart and lungs are normal. The 
abdominal organs are also normal. His legs are remarkable for the 
great forward curving of the tibiae, without any thickness of their 
anterior borders, which are quite ‘sword-like* in sharpness. From 
time to time sharp shooting pain is experienced in the legs. The 
reflexes are normal. Scars of old yaws can be seen on his chest and 
arms. 

Treatment. Potassium iodide in increasing doses, bromides, 
phenacetin, &c., were all apparently useless. 

The chief reason for venturing to bring forward this case is the 
presence of the third tumour. I have been unable to find that such 
has been described before associated with goundou. The associated 
curvature of the tibiae has been noted already by others, but it is not 
referred to in most text books dealing with tropical disease, and the 
same observation largely holds in reference to the continued patency 
of the nasal (lachrymal) ducts. The definite history of yaws preceding 
the appearance of the tumours is also of interest, and the fact that 
nasal discharge was not frequent till after the onset of growth of the 
tumours. 

To what the origin of the third tumour is due I am doubtful. It 
is in a position which would lead one to suspect the cause might lie 
in the Antrum of Highmore, but there is neither apparent thinning of 
bone, discharge from the left nostril, missing or decayed teeth, nor 
depression of the left palate. I could get no evidence to support the 
theory that the nasal passages had been invaded by larvae. 

Goundou has been already reported from Sierra Leone, but I 
believe the disease may be said to be rare in this part of West Africa. 
I regret the patient refused to allow me to operate for the removal 
of any of the tumours, 


ttf 









291 


SUB-DRAINAGE AS APPLIED TO 
THE ANTI-MALARIAL CAMPAIGN 
ON THE ISTHMUS OF PANAMA 


BY 

HENRY SIMMS 

SANITARY ENGINEER, GOLD COAST, FORMERLY CANAL ZONE, PANAMA 

(Received for publication 30 October , igoi) 

With the growing interest at present taken in anti-malarial work, 
and in view of the large works being at present carried out as a result 
of a fuller realisation of its commercial value, it is believed that the 
value of any information on the results of the practical application of 
successful methods not in general use is sufficient warrant for this 
article. It must be remembered that the methods described are the 
outcome of experience with the somewhat unusual tropical climate 
and topography prevailing on the Isthmus of Panama. The rainfall 
is very heavy, averaging about 180 inches on the Atlantic coast and 
80 on the Pacific. The rainy season lasts from May to December, 
inclusive. 

Ordinary methods in the design of sub-drainage for agricultural 
purposes are not applicable. For the agriculturist the object aimed 
at is the removal of subsoil water. For Anopheline extermination 
the object is to remove all water from the surface, and all puddles 
formed by seepage water, however small. All wet places should become 
thoroughly dry at least once every ten days. Here this must be 
accomplished in the face of continuous rains. An engineer without a 
knowledge of the habits of the Anophelines in question (in this case 
Cellia albimanus , C. argyr it arsis , and C. tarsimaculata) would not 
be well equipped for the work. The results obtained by the sub¬ 
drainage work are shown by its effect on the fever rate. This rate 
shows the number of cases sent to the hospitals monthly as a 
percentage of the number of employees appearing on the rolls of the 
Commission. 

At the start of work on the Canal, in the spring of 1904, all the 
energies of the Sanitary Department were directed towards the 
suppression of yellow fever and to the cleaning of the two ports of 



292 


Panama and Colon, the keys to the health of the Isthmus apart from 
malaria. Yellow fever was stamped out towards the end of 1905, and 
the attention of the Department was directed to the reduction of 
malaria. The conditions prevailing were very favourable to the 
propagation of this disease. What drainage had been done had been 
directed towards getting rid of the larger bodies of water. Vegetation 
was rank, and grew close up to the dwelling-quarters. Alongside 
the camps, consisting of unscreened houses, were native settlements, 
78 per cent, of the population of which showed malaria as a result of 
blood examinations in fresh specimens, mostly taken from adult men. 
The place was swarming with Anophelines. White men direct from the 
North, as well as negroes, were placed in the camps. The negroes 
spent their evenings in the native villages. In December, 1905, of 
the total employees 9*63 per cent, were in the hospitals with malaria. 

Systematic measures were immediately started. The general 
scheme was to cut all vegetation growing on soft or soggy places; 
to confine all water in small surface ditches, and to make a copious use 
of crude oil. To this was soon added the screening of buildings and 
cutting of all vegetation for a distance of six or seven hundred feet 
from the houses, which distance seemed to comprise the length of 
flight of the Anophelines. Quinine also played an important 
part. At first the labouring employees would not take the drug, but 
they had no objection to the more palatable mixture of quinine and 
rum subsequently tried. This mixture, giving 4 grains of quinine to 
the wineglass, was made with an inviting colour, and was very 
successful. 

The maintenance of the open ditch system, involving re-grading, 
filling holes scoured out in heavy rains, removal of vegetation and 
algae, application of larvicides, the fact of the collapsing of banks and 
continual inspection, were so expensive, besides being unsatisfactory, 
that the authorities found it necessary to use some more permanent 
methods involving less annual expense in upkeep. They recognised 
that while there was water within reach of mosquitoes there would 
be larvae, as experience had shown that with the greatest care in the 
use of larvicides, and with the best inspection, larvae would escape 
destruction. They reasoned that the only practical method, in view 
of maintenance costs, was to get rid of the water. Sub-drainage was 
considered, and a first shipment of tile was ordered and installed. 



293 

The success of this was so complete that extended sub-drainage was 
decided on. 

Tile drainage work was started early in 1906, but it was not until 
towards the end of the year that sufficient was laid to begin to show 
its effects. During 1907 the work was vigorously conducted. The 
figures given below will show the effect produced. It must be stated 
that the figures given for 1906 do not fairly express the conditions. 
In some districts hospitals had not yet been erected ; also the coloured 
employees had not yet realised the value of the hospitals, and would 
lie about the villages and camps. It was not until a systematic 
inspection was started whereby the District Physician and Sanitary 
Inspector would visit the houses and camps, sending all cases to the 
hospitals, that true statistics became available. The month of March 
is selected as being towards the end of the dry season, and August as 
the period during which the fever rate reaches the highest point. The 
figures show the percentage of employees passing through the 
hospital during the month. The third column shows the annual 
average per 1,000 of deaths from malaria, based on the number of 


deaths in August for the years shown : 

: - 


i 

March. 

August. 

Annual Average per 
1000 of Deaths. 

~ j 

1906 | 

! 

6-32 1 

901 

IO ’55 

1907 

1 

1 

4*16 

5-33 

5*05 

i 9°8 

123 

343 

2*44 


As a rule, the breeding areas giving the greatest trouble here, 
besides small streams, swamps, and low-lying, soggy land, are seeps 
on the sides and at the bases of hills from water following small areas 
of impervious strata. At Ancon this is specially marked. They are 
difficult to find, and the smallest depression holding water can breed 
enough Anophelines to send up the rate for a given camp. 

The tile used was of three sizes.- 4", 6", and 10", porous and 
unglazed. Where the tiles had to receive seepage water only, they 
could be laid at grades as low as one-half per cent., care being 
taken to place a wisp of grass over the joints. If the soil was very 



bad the pipe could be surrounded, and covered for about four inches, 
with coarse gravel or crushed rock, and the trench filled with earth 
other than clay. It is necessary to make systematic borings with a 
6" augur to determine the direction of flow in the soil. Here this 
often follows an almost vertical direction, causing the trenches to be 
very close or very deep. As a rule, however, the main tile follows the 
course of a small stream, with feeders coming in from the side seeps. 
Here the problem is to lay a pipe, as a general rule through stiff clay, 
so that a considerable torrent can pass over it, and such that a few 
hours after it will remove every drop of water and suffer no damage 
from erosion. Such a pipe has of necessity to pass silt in quantities 
during freshets. Experience has shown that a 6" pipe will 
satisfactorily pass silt at a grade of one per cent, and a io" pipe at a 
grade of one-half per cent. These grades can be made ‘ flatter ’ if 
a head can be obtained to act on the pipe, but should be avoided if it 
is possible. Should 4 flatter 1 grades be required, concrete ditches or 
stone ditches set in cement mortar should be used. 

In starting operations on a valley, the first thing to study is the 
best method of straightening the stream with a view to getting greater 
fall and a shorter distance. If the grade exceeds 5 per cent., small 
waterfalls should be introduced to break the velocity over the surface, 
during floods. The pipes should be laid at least 2' 6" below the bed 
of the stream, the trenches should be carefully graded with an 
instrument and the pipes laid evenly and true, with joints open to 
the extent of to They should be firmly imbedded in crushed 
rock on all sides, extending at least 4" above the tile. Rock, broken 
to about 4" cubes, should then be filled in, up to near the surface, and 
the last layer finished off in heavy stone if the scour is great. Some 
small stone must be placed at the top to prevent a too free entry of 
silt in the first rains. All side branches should be treated in the same 
way, and should connect to the main with Y junctions. When 
surface water has to run in volume over the pipe, the outfall should 
be carefully planned so as to pass the water away quickly, and should 
be strong enough to stand the scour that occurs at this point. 

It would appear probable that after a short time the porous stone 
placed over the pipe would completely clog, but this does not 
occur. What happens is that the first heavy rains completely cover 
the stone with about 2" of coarse granular earth. The grass quickly 



2 95 


grows over this and forms a complete mat over which the rain runs 
without erosion. The air from below keeps this mat very porous, 
and the mat then acts as a filter for future rain. Care must be taken 
to cut down any trees the roots of which will interfere with the pipes. 

In the case of seeps, if the borings made do not give sufficient 
data, two or more holes must be dug and the slope that the water is 
following determined. This is sometimes so steep that parallel 
pipes have been laid, io' apart and 5' deep, to catch completely the 
water, as it is essential that all water should be intercepted. 

With regard to cost, it must be remembered that the cost of 
transportation and labour is exceedingly high on the Isthmus. A 
good labourer receives $1.80 per day U.S. currency. Some work 
would be in inaccessible valleys, and complete roads would have to be 
built. Rock would often have to be transported by train and then 
carried over hills. This transportation has made the work 
considerably more costly than it would be in other places. The 
average cost of the tile put in so far is about 35 cents per foot, about 
is. 3d. of which is the first cost of the tile. The cost of maintenance 
of open ditches for the year amounted to 25 cents per foot, so that in 
less than a year and a half the work has paid for itself, apart from, the 
great reduction in malaria and the great saving in the cost of the 
care of the sick. The work is yet far from complete, and much 
remains to be done. 

From the experience gained here it would appear that 
sub-drainage work is the only practical means, taking into 
consideration the cost of upkeep, for permanent anti-malarial work. 
In a climate such as this, and where Anophelines breed in clear 
running water during all months of the year, the annual expense of 
the open ditch system is prohibitive. 




2 97 


A NEW CULICID GENUS 


BY 

F. V. THEOBALD, M.A. 

(Received for publication 6 November , 1908 ) 

Genus Newstcadina, nov. gen. 

Near Orthopodomyia Theobald, but differs in the longer male 
palpi, and the presence of very long scales on the male antennae, and 
also in the wings having Mansonia-Wke scales. 

Head clothed with dense, narrow-curved scales, somewhat 
broadened, numerous upright forked scales broadly expanded 
apically and with flat lateral scales. 

Antennae plumose in the <$, the basal segments with very long, 
narrow, twisted, or wavy scales; pilose in the 9, with narrow 
outstanding flat scales on the two basal segments. Palpi of the $ 
of four segments (?), as long as the proboscis, apical segment very 
small; of the 9 longer than half the proboscis. 

Thorax with rather long, narrow-curved scales ; scutellum with 
rather broad, curved scales; metanotum nude. 

Wings clothed along the veins with large, asymmetrical, flat scales 
(.Mansonia type), and thin, straight, lateral ones beneath them. 
Scaled black and white, giving the wings a markedly ornamental 
appearance. 

Types, aboricollis. D’Emerrez de Charmoy. Ann. Trop. Med. 
and Parasit.* Vol. II, No. 3, p. 257, July, 1900. In the museum of the 
School of Tropical Medicine, Liverpool University. 

Culex fowleri (loc. cit.) The insect described under this name 
is, judging from the single male, a Grabhamia. 

Culex rolandi (loc. cit.) is undoubtedly only Culex micro- 
annulatus , Theo. 


* In the description of this insect in the journal referred to, the following note 
occurred: — ‘ This remarkable insect bears some resemblance to Theobald’s genus 
Lophoceratomyta, though it is quite distinct, and a new genus will probably be erected to 
receive it.— R. Newstead.* 




THE INFLICTED TALIPES- OF THE 

CHINESE 


BY 

FRANK JEANS, M.A., M.B., B.C., Cantab., F.R.C.S, Eng. 

HONORARY ASSISTANT SURGEON, ROYAL INFIRMARY, LIVERPOOL, 

LATE DEMONSTRATOR OF ANATOMY IN THE UNIVERSITY 

(Received for publication 14 November , 1908J 

Actual specimens of the feet of Chinese women are sufficiently 
rare in this country to warrant the following remarks, which are 
relative to a foot kindly presented to the Museum of the Liverpool 
Tropical School of Medicine by Dr. J. Bell, Hong Kong. Since there 
are in the Museum of the College of Surgeons of England, and 
elsewhere, fully dissected specimens, it was considered advisable to 
keep this one in its entirety. It consists of a left foot and the lower 
end of the leg, the section having been made straight across the limb, 
1^ inches above the inner malleolus. The age of the subject is 
unknown, but it is certainly the foot of an adult woman. The 
following are some of the more important measurements : — 

Total length, 5| inches. 

Breadth at metatarso-phalangeal joint, 3 inches. 

Height from the ground to upper surface of scaphoid, 3 inches. 

Maximum height from the ground to lower surface of soft 
tissues, J inch. 

Distance between the supporting points, 3 inches. 

The most striking point at first sight is that the fifth toe is little 
more than half the distance from the heel to the tip of the great toe. 
This description conveys, more than actual measurements, the degree 
of antero-posterior shortening present. But it may be mentioned, for 
the sake of comparison, that normally the fifth toe is situated at a 
distance from the heel of eight-elevenths of the total measurement. 

The position is, roughly, in surgical terms one of Pes Cavus, 
accompanied by a varoid deformity of the outer three toes, and a 
valgoid position of the inner two. The transverse creases often seen 
in talipes acquired from any cause are plainly visible. They run 
horizontally behind the Tendo Achillis, above the heel. The deep 
furrow across the sole divides the foot as a supporting structure into 



two portions -an anterior, triangular, with the base backwards and 
measuring 3 inches from front to back, and a posterior, circular, 
measuring 2 inches in diameter. 

X-ray photographs were kindly taken by Mr. C. Thurston Holland, 
radiographer to the Royal Infirmary, and these throw much light on 
the nature of the deformity and the probable mechanism of its 
production. In the skiagram taken from above, the great toe is seen 
to be in a valgoid position, the greater amount of dislocation being at 
the metatarso-phalangeal joint. This outward position is not more 
than that of the great toes of many European civilized women. The 
second and third toes show the same change at the metatarso¬ 
phalangeal articulations, whilst the interphalangeal joints are acutely 
flexed. In the fourth and fifth toes this flexion of the interphalangeal 
joints completely masks the fact that the first phalanges are pushed 
outwards from their metatarsal bones. 

The first impression, on looking at the side view, is that the 
posterior part of the os calcis is in line with the tibia instead of being 
at right angles to it. Indeed, a radiograph of the os calcis alone, 
when compared with that of a normal bone, explains approximately 
half of the extremely arched condition of the foot. The posterior 
half of the os calcis is set almost at right angles to the anterior half. 
How this alteration in the shape of the calcaneum has been brought 
about is open to question. There are two possible explanations, 
either of which may be sufficient. A consideration of the ossification 
of the bone and of the accompanying radiograph supports the view 
which I take, that the change is mainly in the soft, newly-formed 
portion of the os calcis which is in contact with the so-called 
epiphyseal line (a better term for which would be epiphyseal plane), 
or perhaps in the epiphyseal plane itself. The cartilaginous 
calcaneum at birth contains a small central nucleus of bone which 
ultimately forms the main portion. At about the tenth year a speck 
of bone appears at the posterior end, and these bony formations meet 
at the sixteenth year. According to my view, the pressure, which is as 
a matter of fact applied in an antero-posterior direction, has caused 
the epiphyseal end to slide, as it were, round the main body of the 
bone until it occupies a position below instead of behind it. Against 
this view it may be argued that the epiphysis forms a relatively small 
proportion of the bone, as evidenced by the thin flake of bone which 



can just be separated from the rest of the calcaneum at about the 
sixteenth year, the growth of bone having been actively taking place 
for only six years. But every epiphyseal plane is a travelling plane, 
progressing in a direction away from the centre to the end of the 
bone, and originally this plane must have occupied a position much 
further forward, possibly as far as the point x. 

Another view which occurs to me, but which I regard as less likely, 
is that by the force of the bandages added to that of the Tendo 
Achillis the epiphysis, though forming new bone, cannot form it in 
an antero-posterior direction, and thus perform its true function of 
adding to the length of the bone. It is therefore compelled' to 
deposit ossific material in the direction of least resistance, namely, 
downwards, and this downward prolongation is therefore somewhat 
of the nature of an exostosis. The upward pressure of the ground, 
corresponding to the weight of the child, must on this assumption 
offer less resistance than the bandages. 

In the X-ray photographs it will be noted that, according to the 
former view, the original posterior or epiphyseal end of the 
calcaneum is from y to z y and the patient walks on the insertion of the 
Tendo Achillis, a statement made in 1880, and based on a dissected 
specimen. 

According to the latter explanation, the original posterior end of 
the os calcis is from x to z y and the Tendo Achillis is still inserted 
about half an inch below the point x. 

The os calcis, then, is in any case responsible for the posterior 
half of the arch. In contrast to this, the anterior half is produced by 
a small change in several bones, rather than by a great change in one. 
From the summit of the arch (the astragalus) forwards, there is a 
slight degree of shifting of the articular surfaces of each bone. This 
is a purely anatomical change, and it is doubtful if any particular 
notice would be taken of these individual bones by any anatomist. 

The photographs are of value clinically in that they illustrate, to 
the naked eye at all events, the emphatic statement made long ago 
by Hilton that the fixation of a healthy joint, even for years, produces 
no pathological change. They are, further, of interest to the 
orthopaedic surgeon, in that they shew how much can be done by 
manipulation and continued pressure. 



In the production of the deformity, which takes place during 
the fifth to the seventh years, the size of the foot is reduced by 
bandages which turn the outer toes inwards and under the sole to 
make the foot narrow, while the bandages applied from front to back 
make it short. 

The specimen is regarded by those familiar with the East in years 
gone by as a not excessive instance of the practice. The evidence 
of ulceration seen on the skin is in its slightness quite the exception 
to the rule, gangrene being of fairly common occurrence with the 
frequent loss of one or more of the outer toes. 


DESCRIPTION OF PLATES 

I—X-ray of normal foot from the side. 

II—X-ray of Chinese foot from the side. 

A = axis of anterior half of os calcis. 
P = axis of posterior half of os calcis 

III—X-ray of Chinese foot from above. 

IV, V—Foot from side and from below. 










CONTRIBUTION A L’ETUDE DE 
POROCEPHALUS MONILIFORMIS 


FAR 

A. BRODEN 

DIRECTEUR DU LABORATOIRE 
HI 

J. RODHAIN 

DIRECTEUR DE L’h6pITAL DE.S NOIRS 

Laboratoirc de Leopoldville (Congo beige) 

(Received for publication 16 'November , 1908^ 

Dans une communication precedente, publiee dans ces Annals , 
Vol. I, No. 4, 1908, nous avons relate quelques cas d’infection 
naturelle par larves de Porocephalus moniliformis , Dies; une 
observation chez le negre, quatre observations chez des singes. 

Nous avons pu depuis lors constater d’autres cas d’infection 
naturelle par ce parasite chez le singe et le serpent. Nous avons 
ensuite par une serie d’infections experimentales, 61 ucide complete- 
ment le cycle evolutif de Porocephalus moniliformis. 

I. INFECTIONS NATURELLES 

Malgre les autopsies assez nombreuses de negres originaires de 
differents points du Congo, nous n’avons plus constate d’infection par 
larves de Porocephalus. Comme nous le disions dans notre 
precedente communication, ces infections chez l’homme sont done 
extremement rares. 

Nous avons 6te plus heureux chez les animaux, et depuis notre 
premiere publication, 5 singes et 3 serpents furent trouv^s infectes. 

A. Singes. 

14.X.07: un petit singe Macacus meurt dans la journee; a 
Tautopsie nous trouvons 2 larves de Porocephalus encapsulees dans 
Tepiploon. 

14.III.08: un singe Macacus est trouve mort: dans l’epiploon il 
y a 6 larves de Porocephalus ; pas une seule dans les organes. 



304 


io.IV.o8: petit singe Macacus , mort dans la journee; dans 
lepiploon 2 larves de Porocephalus ; organes indemnes. 

25.IV.08 : singe Macacus mourant, est autopsie immediatement. 
Dans la cavite peritoneale, nous trouvons 1 larve de Porocephalus 
libre ; dans le grand epiploon, 2 larves presque completement sorties 
du kyste, I larve encore bien encapsulee. Pas d’exsudat dans le 
sac peritoneal, pas de lesions organiques. Les larves mesurent 
environ 15 mm. apres fixation ; ce sont 4 9 • 

27.V.08 : singe Macacus est trouve mort au matin. Dans le grand 
epiploon il y a 4 larves de Porocephalus ; dans le mesentere, 1 larve ; 
toutes sont bien encapsulees. Pas de lesions organiques. 

Ces 5 singes, comme les 4 precedents dont nous avons fait mention 
dans la precedente publication, avaient ete achetes a Leopoldville a 
des negres, qui les avaient amenes du Haut-Congo. II nous fut 
impossible de determiner plus exactement le lieu d’origine. 

La disposition des larves de Porocephalus etait identique chez 
tous les singes: nous Tavons decrite dans notre premiere note.* 

Les caracteres morphologiques etaient les memes que ceux 
indiques dans cette meme notice, pour la larve du negre. 

Nous attirons des maintenant T attention sur la grande frequence 
de Tinfection naturelle par Porocephalus chez les singes Macacus au 
Congo. Sur 31 singes que nous avons eus k notre disposition depuis 
fevrier 07 a mai 08, 9 furent trouves infectes. 

Dans l’etude que nous ferons plus loin de Tinfection experimentale, 
nous verrons que ces singes ne se sont pas infestes a notre 
Laboratoire, hypothese que nous avions emise dans notre premiere 
communication. 

B. Serpents. 

A Leopoldville les grands serpents sont relativement rares. Nous 
avons pu en examiner 4, dont 3 renfermaient des Porocephalus 
adultes dans les sacs pulmonaires. 

7.XII.07 : serpent No. 1, Bit is gabonica ,t Su 6 a Leopoldville. 
Renferme dans le sac pulmonaire 29 vers Porocephalus de dimensions 
variables, et 3 vers dans le tissu conjonctif avoisinant les poumons. 

* Ces Annals, Vol. I, No. 4, fevrier 1908. 

f Nous devons la determination des differentes especes de serpents dont il sera question 
dans cette note, & Pobligeance de Mr. Ct. A. Boulenger, du British Museum. Nous Ten 
remercions virement. 



305 

Ces derniers vers sont de dimensions plus petites que ceux renfermes 
dans le poumon. II y avait 15 c?et 159,2 vers furent detruits. 

27.IV.08: serpent No. 2, Python sebae-A nous trouvons dans les 
sacs pulmonaires 13 vers Porocephalus (8 9 , 5 c?), identiques aux 
precedents. 

23.VII.08: serpent No. 3, Python sebae-A renferme dans les sacs 
pulmonaire 17 Porocephalus (10 c?, 6 9 , 1 en tres mauvais £tat). 

Caracteres des Parasites. Ces Porocephalus ont une teinte 
jaune pile, sont de longueur variable; les c? 6tant sensiblement plus 
petits que les 9, et presentant les caracteres generaux decrits pour 
ces parasites. Le nombre des segments est en general de 20 pour 
les 9, et de 17 ou 18 pour les c?. 

Les ceufs , sont arrondis, incolores. A l’interieur d’une premiere 
enveloppe assez mince, se trouve l’oeuf ou l’embryon, entour£ d’une 
membrane epaisse a double .contour, et n’occupant en general que les 
§ ou les | de respace limite par l’enveloppe exterieure. Ce qui 
caracterise surtout Tembryon, c’est Texistence de 4 crochets bifurques, 
paraissant terminer autant d’articles ou de pattes, disposes 
symetriquement. 

Pour obtenir les oeufs en quantite suffisante pour nos essais 
d’infection experimentale nous avons gratt6 les parois des poumons 
chez les serpents infestes, ou dilacere et exprime un ver femelle. 

En parcourant la litterature se rapportant aux Porocephalus , il 
nous a semble qu’il regnait une confusion assez grande dans la 
denomination des differentes especes. L’etude de nos parasites au 
point de vue morphologique et histologique, leur determination exacte 
par comparaison avec d’autres types, necessitant un temps assez 
considerable, nous n’avons pu la faire. Monsieur Gedoelst, professeur 
de parasitologie a l’Ecole de Medecine veterinaire a Bruxelles, a 
bien voulu se charger de ce travail. 

II. INFECTION EXPERIMENTALE 

Pour 61 ucider le cycle evolutif du Porocephalus moniliformis , 
soup^onne mais non encore etudie, il fallait en premier lieu rechercher 
le mode d'infestation de Thote ou des hotes intermediates, ensuite 
reussir l’infection de l’hote definitif, le serpent. 

En effet, toutes les observations recueillies jusqu’a present tant 


x 



chez l’homme que chez les animaux prouvent que le Porocephalus 
moniliformis ne peut arriver k Ye tat adulte que dans les poumons de 
certains serpents. L’homme et les autres animaux ne constituent 
que les hotes intermediates. 

(a) Infection de I'hote intermediate. 

Apres les observations d’infection naturelle recueillies par 
differents observateurs et nous-memes, il paraissait a peu pres certain 
que Tanimal s’infestait* eu avalant accidentellement des neufs de 
Porocephalus. 

Nous avons done recueilli chez les 3 serpents infestes, un nombre 
considerable d’oeufs mis en suspension dans de l’eau physiologique. 
Cette suspension etait ensuite melangee prudemment a du biscuit 011 
de la viande, servant a Talimentation de nos animaux. 

1 0 Singes. 

Singe No /, Macacus .- le 7.XII.07, est infeste avec des oeufs de 
Porocephalus provenant du serpent No. 1 (Bitis gabonica)\ il meurt 
accidentellement le 21.II.08, ou le yje jour apres l’infection. A 
1’autopsie, nous trouvons le grand epiploon farci de jeunes vers 
Porocephalus , tous encapsules. Il n’y a pas de vers libres dans la 
caviti peritoneale, pas de vers dans les organes abdominaux. Dans 
la cage thoracique, il y a 2 vers trbs petits, enkystes a la surface du 
poumon droit.—Ces larves ne montrent encore ni crochets, ni orifices 
sexuels. 

Singe No 2, Macacus : lc 7.XII.07, est infeste avec des oeufs de 
Porocephalus du serpent No. 1 (Bitis gabonica). Ce singe tres vif 
a ete garde en observation durant plus de 5 mois et ne s’est jamais 
montre malade. Il fut tue le 15.V.08, e’est-a-dire le i6ime jour aprfes 
l’infection. 

A lautopsie, le grand epiploon renferme une quantity extreme- 
men t considerable de jeunes vers Porocephalus , tous encapsules; la 
presque totalite des vers se recontre le long des bords libres de 
l’epiploon. 

Le mesentere renferme de nombreux vers enkystes. 

* Nous n’avons pu prendre connaissance, qu’apr&s notre retour en Belgique, de 
Timportant travail de Ch. W. Stiles, Bau u. Entwickelungsgeschichte von Pentastomum 
proboscidium , Rud., u. Pentastomum subcyliridricum. Dies., in Zeitsch. f. wissensehaft . 
Zoologie , LI I, 1891. 




Les ganglions lymphatiques mesenteriques sont hypertrophies et 
renferment plusieurs vers encapsules, a divers stages de developpe- 
ment. 

Le foie, a sa face supero-anterieure, presente 5 vers, tres petits, 
encapsules ; a sa face infero-posterieure, 3 vers. Dans le parenchyme 
de Torgane, il n’y a pas un seul parasite. 

La rate est un peu hypertrophiee, mais ne renferme pas de ver. 

Le diaphragme, a sa face superieure comine a sa face inferieure, 
presente plusieurs vers. 

Les reins ne renferment pas de vers. 

L’estomac presente un ver encapsule, a sa face externe, le long de 
la grande courbure. 

L’intestin grele, a l’union du tiers anterieur avec le tiers moyen, 
presente 1 ver encapsule dans la sous-muqueuse. Dans le contenu 
intestinal nous n’avons pas trouve un seul ver. 

II y a une dizaine de vers encapsules a la face interne de la paroi 
abdominale et du petit bassin. 

Dans la cage thoracique, le poumon gauche est indemne ;—le 
poumon droit, a sa surface, pr6sente 3 vers encapsules, 1 sur le lobe 
superieur, 2 sur le lobe inf6rieur. 

Le pericarde porte 1 ver enkyste a sa face anterieure. 

Un certain nombre de parasites, les plus developpes, ont deja 
des petits crochets et des orifices sexuels. D’autres, moins 
developpes, n’ont pas encore ces appendices. 

Singe No. j , Macacus , est infeste le 7.III.08, avec des oeufs de 

Porocepkalus du serpent No. 1 (Bitis gabonica), gardes dans de l’eau 

physiologique, depuis le 7.XII.07, c’est-a-dire depuis 3 mois. II est 
tue le 1 8 . 11 1.08 ou le I2me jour. A Tautopsie, malgre un examen 
minutieux, nous ne trouvons pas trace de lesions indiquant un 
commencement de developpement des oeufs. 

Singe No. 4, Macacus , est infeste le 7.III.08, avec des oeufs de 

Porocepkalus du serpent No. 1, gardes dans de la terre depuis le 

7.XII.07, c’est-a-dire depuis 3 mois. Les deux premiers mois, la 
terre fut gardee humide, le troisieme mois elle fut negligee, et resta 
plutot seche.—Ce singe est encore en vie. (1.VIII.08.) 

Singe No. 5, Macacus , est infeste le 27.IV.08, avec des oeufs de 
Porocepkalus du serpent No. 2 (Python seba). Est encore en vie 
(1.VIII.08). 



3 °S 


2 C> Rats. 

Le 6.VI.08, 5 rats gris indigenes, sont infesfes avec des oeufs de 
Porocephalus du serpent No. 2, et gardes dans de l’eau physiologique 
depuis le 27.IV.08. 

Rat No. /, tue le 26.VI.08, ou le 21 me jour, pas infecfe. 

Rat No. 2 , tue le 11.VII.08, ou le 36me jour, pas infecfe. 

Rat No. j, trouve mort le 17.VII.08, et a moitie devore par les 
deux survivants. 

Rat No. 4 (Mus rattus), tue el 27.VII.08, ou le 52me jour, est 
infecte. A l’autopsie, nous trouvons dans le grand epiploon de 
nombreuses granulations, grosses comme une petite tete d’epingle. 
L’examen microscopique montre dans ces granulations la presence 
d’un tout petit ver vivant. Le mesentere presente egalement de 
nombreuses granulations. La rate et les reins, portent a leur surface 
quelques granulations identiques ; il n’y en a pas une seule a l’interieur 
de ces organes. 

Les autres organes sont indemnes. 

Rat No. 5 (Mus rattus), tout jeune, tue le 27.VII.08, out le 52me 
jour. A l’autopsie, nous constatons des lesions identiques a celles 
du rat No. 4: les memes organes sont infestes, mais on outre il y a de 
tres rares granulations dans les deux poumons. 

3 ° Coq. 

Un jeune coq fut infeste le 27.IV.08 avec des oeufs de Poroce¬ 
phalus du serpent No. 2 ; il fut tue le 15.VI.08 out le some jour, mais 
n’avait pas contracte d’infection. 

4 0 Canard. 

Un jeune canard C f ut infeste le 27.IV.08 avec des oeufs de 
Porocephalus du serpent No. 2 ; il fut tue le 26.V.08 ou le 3ome jour, 
mais n’avait pas contracte l’infection. 

5° Chacal. 

Un jeune chacal 9 fut infeste le 7.XII.07 avec des oeufs de 
Porocephalus du serpent No. 1 ; il fut tue le 10.IV.08 ou 95me jour, 
mais n’avait pas contracte d’infection. 

6° Chat. 

Un jeune chat sauvage 9 fut infeste le 8.V.08 avec des oeufs de 
Porocephalus du serpent No. 2, gardes dans de l’eau physiologique 



309 

depuis le 27.IV.08. II fut trouve mourant le 28.V.08, ou le 21 me 
jour, et autopsie immediatement 

Dans le grand 6piploon et le mesentere, nous remarquons quelques 
petits points blancs, comme une petite pointe d'6pingle. Au 
microscope, ces petits nodules sont composes d une partie centrale ou 
l'ceuf de Porocephalus plus ou moins modifi6, et d’une partie 
p6ripherique formee par la reaction cellulaire. Dans certains oeufs, 
nous avons retrouv6 encore un ou deux crochets parfaitement 
reconnaissables. 

7 0 Homme . 

Deux hommes et une femme furent infestes avec des oeufs de 
Porocephalus du serpent No. 2. Disons immediatement que tous 
trois etaient arrives au stade ultime de la trypanosomiase, et que chez 
aucun d’eux, Tingestion d’oeufs de Porocephalus ne provoqua le 
moindre symptome pathologique. 

(a) John <3, arrive a la p&riode ultime de la trypanosomiase, 
completement fou; il s’etait refuse energiquement et obstinement k 
tout traitement.—Le 27.IV.08, il avale quelques centimetres cubes 
d’eau contenant des oeufs de Porocephalus du serpent No. 2. 
Ulterieurement il n a pas presente le moindre symptome morbide 
pouvant etre attribu£ aux Porocephalus. Il meurt de trypanosomiase 
le 30.VI.08, c’est-a-dire le 65me jour apres avoir avale les oeufs. 

Autopsie: pas de lesions cutanees, nutrition assez bien conservee. 

Abdomen: dans la cavite peritoneale, pas d’exsudat, gu^re de 
traces de lesions inflammatoires. 

Grand epiploon: dans sa partie superieure pr6sente d’innom 
brables t&ches blanches, opaques, aplaties, ayant jusque 2 mm. de 
diametre. Au microscope, ces taches ne montrent pas ^organisation 
(oeufs de Porocephalus resorbes?). Le long des bords libres de 
Tepiploon, quelques granulations plus ou moins transparentes, grosses 
comme un demi-grain de riz. Il suffit de les dilac£rer legerement et 
prudemment pour en faire sortir un petit ver Porocephalus . 

Intestin grele , sur toute sa longueur pr^sente a la face externe de 
nombreuses granulations, dont la plus grosse ne d6passe pas la moitie 
d’un grain de riz. Quelques-unes ne contiennent pas ou plus de 
Porocephalus t d’autres renferment un jeune ver identique a ceux de 
l^piploon. Incis6 sur toute son 6tendue, Tintestin est vid6 de son 



contenu. La muqueuse presente en de nombreux endroits, des 
petites tumefactions produites par des granulations siegeant dans la 
sous-muqueuse, et renfermant un jeune ver Porocephalus. 

En examinant methodiquement le contenu intestinal recueilli dans 
une cuvette, nous y trouvons I Taenia t quelques Ankylostomes, puis 
une vingtaine de petits vers Porocephalus ; nous en avons retrouve 
encore deux accoles aux replis de la muqueuse. 

Gros intest in: ni a la face exteme ni k la face interne nous 
n’avons trouv£ de granulations; la muqueuse presente quelques 
cicatrices d’ulcerations anciennes. 

La rate et les reins ne presentent rien de special. Les nombreuses 
incisions faites dans ces organes ne revelent pas de Porocephalus. 

Foie: n’est pas augmente. A sa surface, il presente a sa face 
anterieure comme a sa face posterieurc, de nombreuses granulations 
plus ou moins grosses, la plus volumineuse ne depassant pas un demi- 
grain de riz: toutes ces granulations renferment un jeune ver 
Porocephalus. Apres incision, nous constatons que la parenchyme 
hepatique reuferme de nombreuses granulations a ver Porocephalus. 

Cage thoraciquc: les 2 poumons presentent a leur surface quelques 
rares granulations a Porocephalus ; dans le tissu pulmonaire 3 granu¬ 
lations identiques. 

Ganglions lymphatiqucs: nous avons examine 5 ganglions de 
plexus coeliaque, 2 ganglions du mediastin anterieur, sans trouver 
de Porocephalus . 

( b ) Wadi <3, atteinte de trypanosomiase a la periode ultime, 
devenu aveugle apres traitement a l’atoxyl; absorbe le 27.IV.08, un 
peu d’eau avec des oeufs de Porocephalus du serpent No. 2. Depuis 
ce moment, le malade n’a pas presente le moindre symptome morbide 
pouvant etre attribu6 aux Porocephalus. II meurt le 6.VII.08, 
c’est-a-dire le 71 me jour apres l’ingestion des oeufs. 

Autopsie: les constatations sont identiques a celles faites chez le 
sujet precedent, John : infection du grand epiploon, de la tunique 
intestinale, du foie et des poumons. En outre nous avons trouve de 
nombreux ganglions mesenteriques renfermant de 1 k 8 vers Poro¬ 
cephalus k differents stades de developpement. 

Les vers trouves chez ces 2 sujets etaient trcs petits, et ne 
presentaient que 2 a 4 mm. de long. 

(r) Gwangwate 9 , atteinte de trypanosomiase a un stade avanc6, 



incurable; ingere le 27.IV.08 de l’eau avec des oeufs de Potocephalus 
du serpent No. 2. 

Encore en vie le 1.VIII.08. 

En recapitulant brievement ces constatations, nous trouvons que 
1 ’infection experimentale par la bouche au moyen d’oeufs de Poroce¬ 
phalus moniliformis provenant de serpents (Bitis gabonica , Python 
sebae), a reussi chez divers animaux, 2 singes ( Macacus ), 2 rats 
indigenes ( Mus rattus ), 1 jeune chat sauvage, et chez 2 negres. Le 
resultat fut negatif chez le coq, le canard et le chacal. 

Nous ferons remarquer ensuite que chez aucun des animaux 
infectes, nous n’avons pu constater de lesions macroscopiques. Les 
jeunes vers Porocephalus encapsules, etaient fixes sur la sereuse ou 
dans le parenchyme des organes, mais autour d’eux il n’y avait pas de 
reaction inflammatoire. Le Porocephalus , jusqu’ a un certain stade 
de developpement, parait done bien pen irritant pour les organes. 

Nous signalerons ensuite le developpement extremement lent du 
ver. Ainsi chez le singe Macacus No. 2, tue le i6ime jour apres 
l’ingestion des oeufs, les vers avaient au maximum 12 mm. de long, 
la plupart ayant des dimensions moindres. 

Enfin, quant aux organes infestes, nous ferons remarquer que chez 
les divers animaux, singes, rats, chat, pas un seul ver ne fut trouve 
a Tinterieur des organes. Toujours les parasites etaient fixes k une 
sereuse, peritoine ou pleure. Par contre chez Thomme, nous avons 
trouv6 de nombreux vers encapsules dans le foie et quelques-uns 
dans les poumons. 

De ces essais d’infestation experimentale nous pouvons conclure 
que les hotes intermediates s’infestent de Porocephalus monili¬ 
formis , en avalant accidentellement des oeufs provenant de vers 
adultes loges dans les poumons des grands serpents (Bitis gabonica , 
Python sebae). Ces oeufs doivent passer par la trachee et etre avales 
ensuite par le serpent pour arriver a 1‘exterieur par la voie intestinale. 

(b) Infestation de I'hote definitif. 

Pour completer Tetude du cycle evolutif du Porocephalus monili 
for mis, il nous restait a infester un serpent avec de jeunes parasites 
provenant d’un hote intermediate infect6 exp^rimentalement. 

Il nous paraissait probable que dans la nature, les grands serpents 
devaient s’infester en avalant un animal quelconque (singe, rat), 



3 X 2 


porteur de jeunes Porocephalus. Dans les conditions ou nous nous 
trouvions, nous ne pouvions songer a operer avec des serpents boa. 
L*exp6rience fut tentee avec des petits serpents. 

Le 15.V.08, nous faisons avaler a 2 serpents, Causus rhombeatus 
de jeunes vers Porocephalus encapsules dans le grand epiploon du 
singe Macacus No. 1 (infeste experimentalement le 7.XII.07 avec des 
oeufs de Porocephalus du serpent No. 1 ( Python sebae ), et tue le 
i6ie jour apres l’infestation). 

L’un des serpents fut trouve mort le 20.VII.08, et ne put etre 
examine. 

L’autre fut tue le 23.VII.08, ou le 7ome jour apres Tinfestation. 
A Tautopsie nous trouvons dans le sac pulmonaire 4 vers Poroce¬ 
phalus , 2 c? et 2 9 - Ces vers ont une longueur, les S de 2 a 2*5 cm., 
les 9 de 3 a 3*5 cm., apres fixation. Ils sont en tous points identiques 
a ceux que nous avons trouves dans les poumons des grands serpents, 
mais sont de dimensions notablement moindres. 

Ainsi done, nos previsions s’etaint realisees: il est possible 
d’infester des serpents en leur faisant avaler de jeunes vers Poroce¬ 
phalus , et nous pouvons en conclure que dans la nature les grands 
serpents sinfectent en avalant des animaux infestes de jeunes 
Porocephalus. 


CONCLUSIONS 

i° Les parasites que nous avons trouves, comme infection 
naturelle, chez plusieurs singes ( Macacus ) et trois serpents (1 Bit is 
gabonica , 2 Python sebae) t ne sont que deux stades differents du 
meme parasite •. forme jeune chez le singe, forme adulte chez les 
serpents. 

2 0 L’homme et divers animaux, singe, rat, chat, s’infestent en 
avalant accidentellement des oeufs du parasite ; ils ne constituent que 
des hotes intermediates. 

3 0 Les serpents, et surtout les grands serpents, s’infestent en 
avalant un hote intermediate infecte; ils constituent l’hote definitif. 

L’^tude morphologique et histologique des parasites, qui sera 
publiee ulterieurement par le professeur Gedoelst, prouvera si nous 
avons eu raison de considerer, avec Neumann et Looss, le 
Porocephalus constrictus comme la forme jeune du Porocephalus 
moniliformis . 



313 


LITTIiRATURE 

Kn dehors des Auteurs signals dans notre premiere note, nous signalons 
d’apres Neumann (Arch, de Parasitologie, II, 1899):— 

P. M£gnin. Les Parasites et les maladies parasitaires, Paris, 1880. 

C. Wedl. Uber ein Pentastom einer Lowin. Sitrungsberichte der math.-naturw. 

Classe der Kais. Akad. der Wissenschaften, XLVIII, 1 Abth., p. 408, 1863. 

R. Bassi. II Pentastoma moniliforme (Dies.) nella Pantera. II Medico veterinario 
(4)» VI, p. 529, 1877. 

A. Macalister. On two new species of Pentastoma. Proceedings of the Royal 
Irish Academy (2), II, 1874. 

\V. E. Hoyle. On a new species of Pentastomum (P. protelis), from the mesentery 
of Proteles cristatus. Transact. Roy. Soc., Edinburgh, XXXII, 1883. 

A. E. Shipley. An attempt to revive the family ‘ Linguatulidae.' Arch, 
de parasitologie, I, 1898. 

Ch. W. Stiles. Bau und Entwicklungsgeschichte von Pentastotum proboscideum 
Rud. und Pentastomum subcylindricum Dies. Zeitschr. f. wissensch. 
Zoologie, LII, 1891 

C. M. Diesing. Systema helminthum, I, 1850. 

P. J. Van Beneden. Recherches sur 1’organisation et le d^veloppement des 
Linguatules. Ann. des. Sc. Nat. (3), XI, 1849. 

Dans la literature plus recente, nous signalons :— 

Thiroux. Un cas de Pentastomum constrictum observe au S£n6gal. C. R. Soc. 
Biol., LIX, p. 78, 1905. 

Herzog et Hare. Porocephalus constrictus in a native Filipino. Trans. Amer. 
Micr. Soc., XXVII, 1907, p. 41*45. 

Kiewiet de Yonge. Nadere inlichtingen over de Porocephalus moniliformis. 

Geneesk. Zydschr. Nederl. Indie, XLVI, 1906, p. 524. 

Ouwens. Porocephalus moniliformis niet aTleen tot Afrika. Ibid., XLVI, 1906, 
P- 423. 

Waldow. Porocephalus moniliformis bei einem Kamerunneger. Arch. f. Schiffs u. 
Tropenhygiene, Bd. XII, No. 10, Mai 1908. 




A NEW HUMAN NEMATODE 
STRONGTLUS GIBSON I, n. sp. 


BV 

J. W. W. STEPHENS, M.D. Cantab. 

WALTER MVERS LECTl'RER IN TROPICAL MEDICINE, INIVKRSITY OF LIVERPOOL 

(Received for publication 17 November , 1908) 

Some time ago I received from Dr J. Bell, Civil Hospital, Hong 
Kong, some nematodes for diagnosis. The following history 
accompanied them: ‘ They were found by Mr. Adam Gibson, 

Colonial Veterinary Surgeon, in the faeces of a Chinaman employed 
at the Slaughter-house, Hong Kong.’ 

There were ten $ o and nine 9 9 - On examining the males, it 
was evident that they were probably new species of Strongylidae 
from the configuration of the male bursa and the extremely long 
delicate spicules which were extruded in several of the specimens. 
Prof. Eooss, to whom I sent them for examination, wrote: ‘The 
worms you sent arc new to me ; I only know one similar form which 
I found in hares from the neighbourhood of Saw-akin.’ The state of 
preservation was unfortunately very poor, the specimens being much 
distorted, and giving the impression of having been partially dried 
at some time or other. The bursae, however, had preserved their 
contour fairly well, and in one or two specimens it was possible to 
observe the uterine and anal openings in the female. 

The male .—21 mm. long and 0*4 mm. thick towards middle. Head 
attenuated. Two lateral papillae occur, one on either side of the 
buccal orifice. About 0*45 mm. behind the head are situated two 
cervical papillae (fig. 1). 

The Bursa. —Visible to the naked eye is a bilateral appendage. 
Each lobe is a concave-convex lanceolate expansion, the tip of one 
folding over that of the other. The plane of the lobes lies in the 
dorso-ventral plane. Dorsally the edges of the lobe are continued 
forwards, meeting to form a long V-shaped slit, while posteriorly the 
edges run a parallel course until they curve inwards at the tip (fig. 2). 
Ventrally the lobes have, a little in front of their origin, two well- 



marked sub-conical lobules, otherwise the ventral edge is uniformly 
curved from base to apex (fig. 3). 

The disposition of the rays as far as could be seen was as follows : 

Dorsally. An (anterior) ray terminates in a papilla about the 
middle of the dorsal edge ; ( a ) behind this are two (median) rays, the 
posterior of which is the larger, and is curved inwards before it 
reaches the margin. Finally two rays diverging from a common 
stem end in papillae at the tip of the bursa (fig. 4). 

Ventrally. A ray terminates in each of the ventral lobules 
(fig. 5). It was not possible to trace any of these rays from their 
origin from the central mass. 

The distance from the base of the bursa (dorsally) to the tip of 
the folded and curved lobes is about a millimetre. 

Appendages of the genito-anal opening .—Dorsally there is a 
prolongation of the subcuticular layer, which bifurcates posteriorly 
into two appendages which appear to have papillae. Anteriorly this 
is bounded by a crescentic line (fig. (>). Laterally on either side there 
is a small lobule. 

Ventrally there is a conical plate with chitinous margin, and 
closely applied to it dorsally a larger crescentic plate, also with 
chitinous margins, the anterior portion of which is continuous with 
the ventral edge of the bursa (fig. 7). 

The Spicules are about 7 mm. long, thus occupying about one- 
third of the length of the body. In figs. 8 and 9 they are shown in 
different stages of extension. 

The female is about 25 mm. long. The tail end is pointed, and 
in some specimens somewhat curved. The anus is situated 0*2 mm. 
and the uterine opening 0*5 mm. from the tip. The eggs in utero 
measure no x 53 fi (fig. 10). 

The worms are represented in their natural size in fig. 11. 












i 17 


ON THE SUPPOSED OCCURRENCE 
OF FILAR1A IMMITIS IN MAN 

BY 

J. W. W. STEPHENS, M.D. Cantab. 

WALTER MYERS LECTURER IN TROPICAL MEDICINE, UNIVERSITY OF LIVERPOOL 


(Received for publication 17 November , 1908J 

Over and over again the statement occurs in text books of 
medicine and parasitology that this worm has been found by Bowlby 
in man. Not infrequently doubt is thrown on the accuracy of the 
identification of the parasite by Bowlby. On referring recently to 
Bowlby’s communication, I was surprised to find that the worms 
found by Bowlby were Bilharzia and that no mention whatever is 
made of Filaria immitis. Further I found that this fact had 
already been pointed out by Moniez* and an explanation given of 
how the error arose. As, however, this passage in Moniez appears 
to have been overlooked, I think it advisable to call attention once 
more to the facts. 

The following is the report! of the communication made by 
Bowlby to the Pathological Society of London. 

‘ Mr. Bowlby exhibited the urinary organs and parts of other 
viscera removed from two cases of Bilharzia. The first patient was 
an Arab, who was admitted into the hospital at Alexandria under the 
care of Dr. Mackie. The man was suffering from severe cystitis, 
with foul, blood-stained urine. On examination the bladder was 
found to be greatly thickened, and felt as though it contained a 
malignant tumour. In the urine the ova of the bilharzia worm were 
found. Perineal cystotomy was performed to relieve the patient’s 
suffering, but he died a fortnight later. At the post-mortem 
examination numerous female bilharzia worms were found in the 
portal vein, thirty-seven of which, together with the urinary organs 
and portions of the lungs and spleen, had been sent by Dr. Mackie 

* Traite de Parasitologic animale et vej'ctale uppliquee a la medicine, 1896. p. 356. 
t Lancet, 20th April, 1889. p. 78b. 



to Mr. Bowlby. The bladder was contracted and thickened. The 
mucous coat was covered with a shreddy mass of finely fibrillated 
villous growth. The ureters were dilated, with thickened walls, and 
the mucous membrane was covered by a phosphatic deposit. The 
kidneys were in a state of suppuration. Under the microscope the 
thickening of the bladder wall was found to be due to an interstitial 
overgrowth of fibrous tissue, and the mucous membrane had been 
destroyed and was replaced by young fibious tissue. There were 
numerous ova imbedded in the wall of the bladder. The walls of the 
ureters were filled with ova, some of which could be seen in the 
mucous membrane where it had not been destroyed. The kidneys 
showed the changes due to nephritis, and contained numerous ova. 
The lungs were semi-solid; several ova scattered through them. 
The second patient was a boy, seventeen years of age, from whose 
rectum Dr. Mackie removed a tumour (exhibited). The patient had 
suffered from rectal pain and the passage of blood. The tumour 
consisted of a diffuse papillomatous growth, which under the 
microscope was found to consist of a loose, richly cellular, fibrous 
tissue, in the interstices of which were numerous ova. 

‘ Dr. Stephen Mackenzie said the opportunity of studying the 
general pathology of this disease was rare. He asked what was the 
origin of the coagula and fibrous threads often seen in the urine in 
these cases. Were any parent worms found in the bladder? They 
nearly always inhabited the blood-vessels. The ova in the alimentary 
canal were said to have lateral spines, while those in the urinary tract 
had terminal ones.—Dr. Moore had found both lateral and terminal 
spines on ova in both situations.—Mr. Bowlby, in reply, said that the 
process was composed of young fibrous tissue in a state of disintegra¬ 
tion. The parasites were only found in the portal vein.’ 

Then, as already pointed out by Moniez, an abstract with the 
following title appeared in September, 1889, in the Centralblatt fiir 
Bakteriologie, Bd. VI, 1889, P- I 90: — 

‘ Bowlby Mittheilung fiber 2 Falle von Filaria immitis beim 
Menschen (Lancet, Vol. I, No. 16, p. 786). 

‘(1) Bei der Sektion eines Arabers, welcher an Blutharnen 
gelitten hatte und dessen Blasenwand sich bei Lebzeiten schon 
verdickt anffihlte, fanden sich in der Vena portarum zahlreiehe 
weibliche Wiirmer. In der stark verdickten Blasenwand waren 



zahlreiche Eier eingebettet. Auch in den Harnleitern und Nieren 
fanden sie sich, sowie, in geringer Zahl, in den etwas derb 
anzufiihlenden Lungen. (2) Bei einem 17 jahringen Knaben wurde 
ein Tumor im Rectum entfernt. Derselbe erwies sich als aus einem 
lockeren reichlich zellenhaltigen, fasrigen Gewebe bestehendm in 
dessen Zwischenraumen zahlreiche Eier lagen.’ Kurth (Berlin). 

This accidental or erroneous use of the name Filaria immitis for 
the worms, which as the original shows were Bilharzia, has probably 
been the source of all the following erroneous misquotations. At 
any rate it is clear that in these cases there is no question of Filaria 
immitis but of Bilharzia. 

To make quite certain of this I wrote to Mr. Bowlby, and the 
following is an extract from his reply to my letter :—‘ I did not know 
before that I was supposed to be the discoverer of this Filaria. I 
am quite innocent of any knowledge of the said parasite, and the 
paper you refer to was on some cases of Bilharzia. If you can correct 
the error, please do.' This then disposes of Bowlby’s supposed 
record. 

Finally there is the record by Braun* of the supposed occurrence 
of F. immitis in man. Braun’s words are the following :— 1 Ich fuhre 
hierbei dass im Jahre 1885, in Dorpat in der leiche eines Russen die 
zu Praparieriibungen benutzt wurde sehr lange Nematoden in 
grosserer Zahl in den Venen gefunden worden sind ; ich habe die 
wohl-erhaltenen Wiirmer selbst gesehen und korserviert; an ihren 
Filarien natur ist nicht zu zweifeln, jedoch bin ich nicht imstande 
mehre auszusagen, da ich die Parasiten nicht mehr untersuchen 
konnte.’ 

In this case, however, as the species was not identified, I do not 
think we are at present justified in including F. immitis among the 
parasites of man. 


* Braun, Max. Die Tierischen Paiasiten <k*s Menschen, Vierte AuHage, p. 295. 





A NEW POROCEPHALUS 
[POROCEPHALUS CERCOPITHECI 

n. sp.) 

BY 

ANTON BREINL and EDWARD HINDLE 

From the Runcorn Research Laboratories of the Liverpool School 

of Tropical Medicine 

(Received for publication g December , 1908,) 

At the autopsy on one of our experimental monkeys (a large 
female Cercopithecus callitrichus ), in the left lower lobe of the lung, 
a subpleural cyst about 3 mm. in diameter was noticed. This cyst 
contained an immature example of a Porocephalus coiled up inside it. 
Although the whole lung was carefully dissected, no other specimens 
were found; the other organs and the gut were also searched 
without success. 



Kig. i. 

The body is a uniform greyish white in colour (fig. 1). It 
exhibits a separation into head (cephalo-thorax ?) and body, the 
latter being divided into about 45 annuli, which become very 
indistinct towards the posterior end. The distance between the 
annuli is greatest towards the anterior end, and gradually diminishes 
posteriorly. 

The dimensions of this specimen are :—Length, 10 mm.; diameter, 
at anterior extremity, r 17 mm.; and diameter at posterior extremity, 
ergo mm. 








322 


The short, bluntly-rounded head (fig. 2) is cut off from the body by 
a distinct groove passing completely round the animal. The dorsal 
surface of the head is slightly curved. The ventral surface is convex 
and bears two pairs of hooks, one on each side of the median mouth. 
This latter is surrounded by a chitmous ring, oval in shape, about 
019 mm. in length, and 0*8 mm. in breadth. The inner hooks are 
single, each consisting of a stout, strongly-curved chitinous process 
c24 mm. in length. In the outer pair, from the base of each hook 
arises a slightly curved unciform appendage (o*n mm. in length), as 
in all the hooks of Porocephalus najae-sputatricis , Leuckart. The 
hooks of both the outer and inner parts are jointed at their bases. 

The body exhibits a median line running along the ventral surface 
to the posterior extremity. 



The anus is a small aperture situated at the apex of a small 
papilla at the posterior extremity. Immediately beneath it is the 
genital aperture. 

This species is distinguished from the known species of 
Porocephalus by the presence of an appendage on the outer pair of 
hooks only. We propose the name Porocephalus cercopitheci in view 
of the host. 

The type specimen of this species is in the museum of the 
Liverpool School of Tropical Medicine. 



323 


COMPARATIVE 

CHEMO-THERAPEUTICAL STUDY OF 
ATOXYL AND TRYPANOCIDES 
PART II 

BY 

M. NIERENSTEIN, Ph.D, 

J. W. GARRETT INTERNATIONAL FELLOW AND RESEARCH DEMONSTRATOR, 
LIVERPOOL SCHOOL OF TROPICAL MEDICINE 

Trom the Runcorn Research Laboratories of the Liverpool School 

of Tropical Medicine 

(Received for publication 9 December , 1908; 

Our previous work 1 on the chemo-therapeutics of Atoxyl has led 
us to the conclusion that a combination in vitro takes place between 
proteins, and Atoxyl mono-acetylated Atoxyl and mono-benzoylated 
Atoxyl respectively; whilst, on the other hand, such a combination 
does not occur between proteins, and sodium arsenate acetyl-benzoyl 
Atoxyl and sodium-p-hydroxy-phenyl-arsenate. This work has been 
continued by injecting the above-mentioned drugs into experimental 
animals ; these reacting in an analogous way to the serum-proteins 
in vitro , with only one exception—acetyl-benzoyl Atoxyl—which 
combined with the serum proteins in vivo. This reaction, however, 
was only to be expected, as the organism saponifies the acetyl group, 
and the resulting benzoyl Atoxyl acts in the same way as mono¬ 
benzoyl Atoxyl in vitro. 

Technique. —Rabbits were injected for several months, twice 
weekly, with Atoxyl, Sodium arsenate, acetylated and benzoylated 
Atoxyl, Benzoyl-acetyl Atoxyl, and Sodium-hydroxyl-phenyl-arsenate. 
Usually 1 c.c. of 1 per cent, solution of the drug was injected. 

After a time, 20 c.c. of blood was taken from the jugular vein, and 
the serum used for analysis. 

The arsenic was estimated in the same way as in our previous 
work; the slightly modified Sanger’s method being adopted. Gold 
chloride was used as developer in preference to hydrochloric acid. 

The results of the experiments are given in the following table 



324 


(Table I). For comparison, our previously recorded 
in vitro are appended : — 

observations 


Table 

I 



Drug Chemical Constitution 

Arsenic 

in vivo 

Estimation 

in vitro 

Number of 
Experiments 
in vivo 

Atoxyl 

1 

As0^^ a 

0 

NH, 

arsenic 

present 

arsenic 

present 

5 

Sodium arsenate 

— 

arsenic* 

absent 

arsenic 

absent 

5 

^K8S* 

Acetylated Atoxvl | | 

\/ 

N< ^CO • CH„ 

arsenic 

present 

arsenic 

present 

2 

Benzoylated Atoxyl | 

\/ 

N <CO • C„H 

arsenic 

present 

arsenic 

present 

2 

Benzoyl-acetyl 1 

Atoxyl 

*-3S‘ 

0 

N /CO • CH, 
\CO • C,;H- 

arsenic 

present 

arsenic 

absent 

2 


Sodium-p-hydroxyl- 

phenyl-arsenate 



arsenic arsenic 

absent absent 


* In one of these experiments arsenic was found to be present. The fact that 
this particular serum contained haemoglobin may explain the exception. 



325 


Similar experiments were carried out on two donkeys in order to 
obtain larger quantities of blood, so as to make a more detailed 
examination of the distribution of the arsenic with regard to the 
constituents of the blood. 

This table shows that the haemoglobin contained arsenic in both 
cases: — 

Table II 

Atoxyl Sodium Arsenate 

Haemoglobin arsenic present arsenic present 

Stroma arsenic present arsenic absent 

Serum arsenic present arsenic absent 

The above recorded experiments confirm and extend the view 
that the amido group in Atoxyl and allied compounds in vitro , as 
well as in vivo , combines with the serum proteins. 

After the mode of the combination of Atoxyl and serum proteins 
in the animal organism had been established, it seemed necessary to 
estimate the amount of Atoxyl which is secreted in order to form an 
idea as to how much of the drug is actually left in the body. 

For this purpose a horse was injected subcutaneously with Atoxyl, 
and the urine and faeces analysed. The arsenic was estimated 
according to Dupas-Gilier’s 2 iodine method, using Gileas’s and 
Shearer’s modification. 

Table III gives the amount of Atoxyl injected and recovered in 
urine and faeces : — 

Table III 

Urine 


Date of Injection 

Amount Injected 

Date of Collecting 

Arsenic R 

28.2.08 

i gm. Atoxyl 

1.3.08 

81% 

2.3.08 

1 gm. Atoxyl 

3-3-o8 

83% 

5-3-08 

1 gm. Atoxyl 

6.3.08 

78% 

7.3.08 

1 gm. Atoxyl 

8.3.08 

82% 

10.3.08 

2 gm. Atoxyl 

11.3.08 

79 % 

12.3.08 

2 gm. Atoxyl 

13.3.08 

85% 

16.3.08 

2 gm. Atoxyl 

17.3.08 

82% 

19.3.08 

2 gm. Atoxyl 

20.3.08 

80% 

22.3.08 

1 gm. Atoxyl 

23.3.08 

86% 



326 


Faeces 


Date of Injection 

Amount Injected 

Date of Collecting 

Arsenic Recovered 

12.3.08 

2 gm. Atoxyl 

13.3.08 

4 % 

16.3.08 

2 gm. Atoxyl 

17.3.08 

2% 

19.3.08 

2 gm. Atoxyl 

20.3.08 

5 % 


The chemical details of this work will be published shortly. 
Atoxyl has been found to be secreted in the urine, as: — 


(1) p-amino-phenyl-arsenious acid; 

(2) p-oxy-phenyl-arsenious acid; 

(3) Arsyl-oxy-carbonyl. 

The presence of this third compound in the urine is of practical 
interest, in so far as its formation can only be explained by assuming 
that Atoxyl is acetylated in the organism, and afterwards the 
p-amino-phenyl-arsenious acid is transformed into Arsyl-oxy- 
carbonyl. Similar observations 3 have been made on the secretion 
of p-toluidine, in which case Methyl-oxy-carbonyl is formed. It is 
quite possible that this may explain the fact that acetylated Atoxyl 
is less toxic than Atoxyl. 

Besides arsenic, anilin was found to be present, only in the faeces. 
The faeces were treated with alkali, and steam passed through the 
mixture. The condensed steam gave distinctly, with bleaching 
powder, Hoffmann’s reaction. 

With regard to the distribution of Atoxyl in the organism, it has 
been shown by different authors that arsenic is deposited in all organs 
to a greater or lesser extent. J. Magalhaes 4 states that Atoxyl does 
not permeate the meninges. We have attempted to verify his 
statement by using the cerebro-spinal fluid of infected donkeys 
withdrawn at different intervals after the last administration of the 
drug. 



3-7 


Table IV gives the results of the analysis : — 


Table IV 


Days elapsed since last Injection 

7 days 

1 day 
9 days 

2 days 

5 days 
9 days 

3 days 

6 days 


Arsenic Examination 

arsenic present 
arsenic absent 
arsenic absent 
arsenic present 
arsenic present 
arsenic absent 
arsenic present 
arsenic present 


These experiments prove conclusively that the meninges are 
permeable to Atoxyl, as in the majority of the recorded cases arsenic 
could be detected in the cerebro-spinal fluid. 


LITERATURE 

1. M. Nierenstein. Comparative Chemo-therapeutical study of Atoxyl and 

Trypanocides, Part I. Annals of Tropical Medicine and Parasitology, Vol. IT, 
No. 3, p. 249. 

2. Zeitschrift fur angewandte Chemie, Vol. 22, p. 378. 

3. See Hoppe-Seyler’s Zeitschrift fur physiologische Chemie, Vol. 12, p. 295, also 

S. Frankel, Die Arzneimittel-Synthese auf Grundlage der Beziehungen 
zwischen chemischen Aufbau und Wirkung, 1906, p. 179 

4. Jos£ de Magalhaes. Etude au point de vue therapeutique de la permeabilite 

mening£e dans la Trypanosomiase humaine. XV Congr&s International de 
Medicine, Lisbonne, 1906. Fascicule 2, p. 304 ; see also Breinl and Todd, 
British Medical Journal, 1907, January 19th, p. 132. 




329 


CHEMICAL NOTES ON ATOXYL 


BY 

M. NIERENSTEIN, Ph.D. 

From the Runcorn Research Laboratories of the Liverpool School 

of Tropical Medicine 

(Received for publicatioti g December , 1908 ) 

The following is a record of chemical observations made during 
the last two and a half years whilst working on the chemical 
constitution and physiological action of Atoxyl. 

Water of crystallisation .—The differences in the amount of 
arsenic found in samples of Atoxyl due to the differences in the water 
of crystallisation (Moore, Nierenstein and Todd, 1 Ehrlich and 
Bertheim, 2 and others), suggested an exact estimation of the water 
of crystallisation and * adhering moisture.’ Experiments have proved 
that Atoxyl contains from one half to one and a half molecules of 
adhering moisture, and exactly two molecules of water of crystallisa¬ 
tion. Further observations showed that Atoxyl loses the adhering 
moisture in about five weeks when standing over concentrated 
sulphuric acid ; after this time no further loss of weight could be noted 
in the course of six months. It contains, then, two molecules of water 
of crystallisation which are only lost on drying for three hours at 
160 0 C. Atoxyl, after having been kept over sulphuric acid for at 
least five weeks, has, therefore, the formula: — 

yOH 

H a N * C G U i • AsO^Q^ a + 2 H tJ 0 , corresponding to 23*88% arsenic. 

Therefore, in order to administer always the same amount of arsenic 
in a given dose of Atoxyl, it is advisable that Atoxyl should be kept 
in a dark desiccator over concentrated sulphuric acid for at least five 
weeks before use. The dark desiccator ought to be used on account 
of the decomposition of Atoxyl solutions when exposed to light for 
some time. 

Inorganic arsenic .—Two distinct brands of Atoxyl manufactured 
by the Vereinigte Chemische Werke, Charlottenberg, have been 



330 


supplied to us since 1905 ; one having the appearance of a white 
powder, the other of distinct crystals. It was found that the powder 
contained free inorganic arsenic (from 0*4 to 0*9%), which is easily 
detected by passing H 2 S into the slightly acidified solution. The 
crystalline Atoxyl did not contain any inorganic arsenic. 

Parafuchsin .—On two occasions it was noticed that the freshly 
prepared solution of Atoxyl was of a red colour; this was due to the 
presence of parafuchsin, which is formed as a by-product during the 
preparation of the drug. 

Yellow Atoxyl .—One special supply of Atoxyl sent out to Uganda 
gave rise, in the hands of Captain A. C. H. Gray, 3 on injection, to 
violent toxic symptoms and blindness. The qualitative examination 
of the drug showed that it dissolved in strong alkali (40% KOH) with 
a yellow colour; this colour reaction has never been observed by us 
in any other sample of Atoxyl. This Atoxyl contained free 
inorganic arsenic, traces of free anilin, and a second substance which 
is probably an oxidation product of Atoxyl. It is quite possible that 
this substance may be responsible for the untoward effects. 

Therefore, it would seem advisable that Atoxyl should be tested 
with strong alkali before use. 1 c.c. of a 5% solution should be mixed 
with 2 c.c. of strong alkali and left standing for a few minutes, and if 
the solution shows a yellow coloration the Atoxyl should not be used 
for treatment. 

A full chemical report on the ‘ yellow Atoxyl ’ has previously been 
published. 3 


LITERATURE 

1. B. Moore, Nierenstein and Todd. Bio-chemical Journal, Vol. II, 1907, p. 300. 

2. Ehrlich and Bertheim. Berichte d. dcutscken chemischen GeseUschaft, 

Vol. 40, p. 3296 

3. A. C. H. Gray. Quarterly Report on the Progress of Sleeping Sickness and 

Medical Treatment of Sleeping Sickness in Uganda, 1908, pp. 32 and 35. 



NOTE SUR LE ROLE DES TABA- 
NIDES DANS LA PROPAGATION 
DES TRYPANOSOMIASES 


PAR 

Le I)R. EDMOND SERGENT 
(Received for publication 25 January , 1909) 

Recemment plusieurs auteurs anglais ont bien voulu rapeller les 
experiences que nous avons faites sur la transmission des trypano¬ 
somiases par les insectes piqueurs. Mais leurs citations contiennent 
parfois une erreur, involontaire, et qui consiste a nous faire dire que 
nous avons reussi a donner le nagana avec des Taons qui avaient 
pique 48 heures auparavant un animal infecte.* L'erreur provient 
de ce que ces auteurs ont du se documenter sur le tableau page 679 
du tome XX des Annales de I'Institut Pasteur , tableau dans lequel 
s’est glissee une erreur de typographic faisant attribuer au nagana 
ce qui revient au debab. Cette erreur a ete corrigee dans un erratum 
publie page 880 du meme tome XX des Annales de I'Institut Pasteur , 
erratum qui a evidemment echappe aux auteurs en question. 

Comme cette question de la transmission des trypanosomiases 
est a l’ordre du jour je crois utile de rappeler ce qu’ont 6te nos 
experiences relatives au debab, que Ton trouvera in-extenso dans les 
Annales de I'Institut Pasteur du 20 janvier 1905 (T. XIX), pages 
31 k 41. 

Nous avons montre que les deux especes de Taons le plus 
repandues dans le Tell algerien: Atylotus tomentosus et Atylotus 
nemoralis peuvent transmettre' exp6rimentalement Tinfection d’un 
animal malade a un animal sain quand les piqures le suivent 
immediatement. Dans une experience nous avons etabli que la 
piqfire de Taons ayant suce le sang d’un animal malade peut infecter 
un animal sain encore apres 22 heures d’intervalle entre les deux 
piqures. 

• Dutton, Todd et Hanin^ton. ‘ Annals of Trop. Med. and Parasit.,’ Vol. I, 2nd note, 
j). 212. 

Kinghorn et Montgomery. ‘ Annals of Trop. Med. and Parasit.,* Vol. II, p. 86. 




333 


ON THE NOMENCLATURE OF THE MAM¬ 
MALIAN TRYPANOSOMES OBSERVED IN 
NORTH WESTERN RHODESIA 

BY 

R. EUSTACE MONTGOMERY, M.R.C.V.S, 

AND 

ALLAN KINGHORN, M.B. Toronto 

JOHNSTON COLON IA1. FELLOW, INI YKRSITY OF LIVERPOOL 


Of the Expedition of the Liverpool School of Tropical Medicine to the 

Zambesi, 1907-1909 

(Received for publication 15 December , 1908 ^ 

In view of the conclusions at which we arrived as to the mature of 
the trypanosomes found in bovines of North Western Rhodesia, it 
appears desirable to briefly outline the subject from the standpoint we 
have taken. 1 We would observe that at the time our report was 
despatched from the Luapula we had before us, with small exception, 
only literature bearing date prior to April, 1907, and on the evidence 
therein contained, we did not feel justified in discussing the question 
of the morphology or specificity of our trypanosomes more fully than 
was done, and it will no doubt have seemed to one noting only the 
date of publication as though our duty had been somewhat neglected, 
and our conclusions based in defiance of or without due regard to the 
results of those workers whose communications had been public 
property in Europe for upwards of fifteen months prior to the 
appearance of our report. Had we before us at the present moment 
only the evidence already advanced, we would hesitate to re-open the 
question ; but the perusal of the literature now with us, and more 
especially our own more recent observations on trypanosomiasis in 
other areas of Northern Rhodesia, justify, we consider, this further 
exposition. In a forthcoming report we shall have occasion to refer 
to and to compare with that on trypanosomiasis in North Western 
Rhodesia; and since we purpose utilising this as a basis for critical 
comparison, it is highly desirable that our position and views 
z 



334 


concerning these trypanosomes, to which we have referred as 
T. dimorphon and T. vivax , should be most clearly stated ; so that, 
whatever be the faults in our deductions, these may be recognised 
and corrected by those to whom the necessary facilities are available. 
Owing to our protracted absence from Europe and the relative paucity 
and delay in the receipt of current scientific literature, we shall be 
precise in specifying all communications referred to, since it is not 
unlikely that at the time of writing important papers bearing on this 
particular case may have appeared in print, but will remain inaccessible 
to us for some months. 

Probably no sub-division of the Protozoa presents more difficulties 
in the way of classification than does the genus Trypatiosoma Gruby, 
emended by Laveran and Mesnil in igoi so as to exclude those 
organisms of fish which carry both an anterior and a posterior 
flagellum. On morphological grounds Liihe has created the genus 
Trypanozoon to contain only those trypanosomes parasitic in the 
blood of Mammals; beyond this no further attempt has been made to 
classify or arrange the genus; and, indeed, any additional sub¬ 
division would be impracticable to a Zoologist. We are in a transition 
stage, when rumours of multiplication cycles, resting stages and forms 
which from their appearance are unrecognisable, even as flagellates, 
are abroad. At the present time, we have positive knowledge only of 
the well-known forms found free-swimming in the blood; and these 
presenting such close analogies to one another, offer no opportunity to 
the Protozoologist of satisfactorily adopting any further classification 
based on morphological features. It is possible that the more exact 
and more strictly cytological technique which is now being employed 
in the study of these organisms may bring to light some new features 
in their structure to serve as a fixed point from which morphological 
classification can commence; but until it has been employed 
uniformly on all known species, and critical comparisons made, it is of 
little more value than the older methods used both by Zoologists and 
Pathologists. 

Any classification dependent upon a variable feature and allowing 
margins to the personal equation must be prone to error, and cannot 
be unquestionably accepted. Yet we find that Zoologists and 
Pathologists alike accept as species forms of trypanosomes 
distinguished from others on morphological grounds. T. dvnorphon 



335 


is admitted to be a ‘ good species/ and solely from its morphology ; 
T. equiniim and T. theileri , the former more especially, are also 
constituted largely from their unique appearance in the blood. 

A clinician may subdivide the diseases due to trypanosomes on 
symptomatological or epidemiological grounds: and a pathologist 
may add to these the results of his observations on animals experi¬ 
mentally infected Here again absolute distinction between any two 
species may be impossible, even though it be known that the 
morphology of each is markedly different. Thus the experimental 
disease, or the 4 animal reactions * due to T. evansi and T. dimorphon , 
show very little variation in most Mammals; yet these organisms are 
clearly distinguishable on morphological grounds. Again, it would 
be a matter of the utmost difficulty to state specifically that 
T. gambiense was not the same as T. evansi , if only dead organisms 
were available. But if experimental animal reactions are considered, 
and the natural disease due to each be observed, they can be most 
strikingly separated. 

However lacking in conformity to Zoological rules, and however 
imperfect and crude, it is possible, by adopting a combination of the 
results given us by morphological and pathological studies, to 
constitute various groups of mammalian trypanosomes, which to a 
worker removed from such facilities as exist in the research centres of 
Europe are, to our mind, helpful. An observer in the field may place 
a detected trypanosome, tentatively, in one of the classes with a 
minimum amount of work, and since he will be in possession of only 
a modicum of research equipment and a limited stock of experimental 
animals, he will be in a position to utilise these to their best advantage, 
and by adopting a certain degree of uniformity can bring his 
observations into line with all work previously carried out on the 
class of organisms to which his own approximates. Absolute 
differentiation of closely related trypanosomes is quite impossible to 
such a worker : a rigorous and critical morphological comparison must 
be effected, and final diagnosis will rest upon the result of this and 
that of ‘ cross-inoculation * into animals believed to be immunised 
against the organisms with which comparison is being made. 

For such work ‘ types ’ of known and approved origin must be at 
hand, as well as the necessary animals for cross-inoculation. This 
must remain the work of an acknowledged centre T assuming the 



336 

functions of a museum wherein all type species are maintained and 
strictly guarded. 

If we eliminate T . lewisi and allied species as not occurring in the 
blood of the higher mammals, there remain thirteen named species 
all more or less accepted and distinctive, which an observer may meet 
in man or the domestic animals. These are T. gambiense , T. evansi , 
T. brucei , T. equiperdum , T. equinum, T. dimorphon, T. theileri 
(including some of Lingard’s forms met in India), T. vivax , T . nanum , 
T. congolense , T . cazalboui , T. pecaudi , and 7 \ sudanense . On 
morphological grounds it is possible to distinguish: — 

T. theileri , on account of its relatively immense size, and further 
from its animal reactions, being parasitic only in the blood of 
bovines. Dutton and Todd, however, describe a trypano¬ 
some morphologically recalling this organism as occurring in 
the blood of a Tragelaphus scriptus in the Congo Free 
State. 2 

T. equinum , which in stained preparation (dry method) shows a 
uniquely small blepharoplast. On epidemiological grounds 
in addition; by being limited to South America. 

By adopting certain standards of animal reaction, we can regard as 
distinctive : — 

T. gambiense , as being the only one, so far as known, pathogenic 
to man. The human subject not being available for inocu¬ 
lation, the diagnosis of forms recalling this species morpho¬ 
logically would remain work for comparison with types. 

T. equiperdum , on the grounds of production in equines of quite 
characteristic clinical symptoms; and being unique among 
trypanosomiases of the lower animals in transference 
naturally by coitus. 

Nine named species remain to be dealt with. By the adoption of 
morphological and animal reaction standards these can be sub¬ 
divided into three groups, having as their types, in point of view of 
priority, T. evansi , T. dimorphon , and T. nanum , respectively. 

The standards we adopt are arbitrary, and are open to criticism, 
as any must be which depend upon more or less relative features of 
size and pathogenicity. 



Considerable variations are noticeable in the appearance of 
individual trypanosomes of any strain: we have the so-Called ‘ male ’ 
and ‘ female ’ elements; and gross measurements are found to vary. 
But, be it noted, these variations are within limits. It would be most 
exceptional to find, for instance, any T . evansi of less than 20 n or of 
more than 35/1 in length; and despite the tens of generations which 
many trypanosome strains have produced in various laboratories, we 
find the morphological features remain practically constant. There is 
not that tendency to develop new types, to revert to an old form, or 
otherwise to vary, which has more than once been suggested. We 
may with comparative safety, then, group into one class some of 
those nine species which conform approximately to the dimensions of 
T. evansi. Disregarding the differences in ‘ male, ‘ female ’ and 
‘ indifferent * forms, these species would be monomorphic and of 
relatively large size. 

On the other hand, there is a group of three species the morpho¬ 
logical appearances of which differ markedly. In the course of 
infection by T. dimorphon y forms closely resembling T . evansi are 
encountered, but at the same time, or in the same animal, or, capable 
of production in another animal, trypanosomes markedly smaller, 
measuring only 10 fi to 15 ju in length, and without any free flagellum, 
make their appearance. It has been shown that these are but various 
manifestations of the same organism, which is consequently far from 
being monomorphic. 

Of the nine species to which we refer, six show similar reactions 
towards experimental laboratory animals; three differ markedly. 
The value of animal reactions may be disputed, and when limited in 
amount, results are apt to cause confusion. Particularly is this the 
case with the donkey, cattle, sheep and goats, largely, we think, 
owing to the multitude of races and breeds employed, and also because 
many of them are country-bred animals whose ancestors have 
probably been exposed for generations to trypanosome infections. 
These animals, too, show a degree of individual idiosyncrasy which is 
prone to give rise to error. Monkeys, especially Cercopithecus and 
rhesus , domestic rabbits, guinea-pigs, white rats and tame mice, and 
to a slightly less degree, dogs of both European and native blood, are, 
however, fairly constant in their susceptibility or their insusceptibility 
to any one strain of trypanosome. During the many generations 



338 


which have been maintained of such organisms as T . gambiense , 
T. evansi and T. dimorphon in all the usual laboratory animals, there 
has never been any suggestion that a new type has been created. 
Increased and decreased virulence may and does occur, but this is less 
marked, under normal maintenance of the strain, than was hitherto 
supposed, and there is always a tendency to reversion towards the 
original state. 

By means of these two factors the nine remaining trypanosomes 
can be conveniently grouped as follows: — 

A. Trypanosomes, pathogenic towards most domestic animals, 

and producing a rapidly fatal infection in the usual 
laboratory animals: 

(a) monomorphic, of large and fairly constant size, from 
20 fi to 35 n in length, and carrying a distinct free 
flagellum. Type— T. evansi. 

([b ) di- or polymorphic, of very variable size, occurring in 
at least two forms: (i) ‘short/ measuring from io/k 
to 15>u in length, and devoid of a free flagellum; 
(2) ‘ long/ which may attain 35/1, and bearing a 
variable length of flagellum. Intermediate forms 
also occur. Type— T. dimorphon. 

B. Trypanosomes, pathogenic to certain domestic animals, and 

without apparent effect when inoculated into the usual 
laboratory animals (monkey, dog, rabbit, guinea-pig, rat and 
mouse). An imperfectly studied group, which includes 
T. nanum , T . vivax and T. cazalboui . 

We consider this grouping to be sufficiently exact for the 
temporary purposes of a worker in the Tropics. Unless the organism 
with which he is dealing shows any striking peculiarity in either 
morphology or animal reaction, he can do little more than assign it 
to one of these groups. 

In North Western Rhodesia we isolated from cattle, sheep and 
dogs three trypanosomes, one approximating to each of the groups, 
and we referred to these as T. brucei , T . dimorphon and T. vivax for 
the reasons set out below. 



339 

*1. T. evansi group, which includes T. brucei and 2 . sudanense? 

The validity of T. brucei , save on the grounds of cross-inoculation* 
is open to question, unless it be pre-surmised that each species of 
trypanosome has its own particular genus or species of biting fly to 
bring about dissemination. In Africa, where both T. brucei and 
T. evansi occur, it has become usual tp refer to Glossina and 
Tabanidae as the respective transmitters. In the same manner an 
evansi- like organism, which in nature is supposedly spread by 
Glossina , or which is met with in cases of ‘ tsetse-fly disease/ is 
referred to as Trypanosoma brucei. This, we take it, implies the 
acceptance of T. brucei as the local type of the evansi group, and we 
consequently assigned to it our dog trypanosome. But it is no more 
possible to distinguish between T. evansi and T. brucei in the field, 
unless the supposed transmitting factor be considered, than it is to 
differentiate T. evansi and T . sudanense . 

2. T. dimorphon group, including T . congolense 4 and T. pecaudi? 

In their action on laboratory animals these three species coincide 
very closely; and in their morphology, as shown in stained film (dry 
method), T. congolense is almost identical with the T . dimorphon at 
Paris. Writing on the subject of these two, Martin, Leboeuf and 
Roubaud 5 hold that in the French Congo the latter is more active, 
moving more readily across the field and producing more extensive 
lateral displacement of the corpuscles; and they contend that a little 
experience will enable an observer to distinguish between the two. 
Assuming that these two species are distinct, based on the strains in 
Europe, it is not to be forgotten that in countries such as parts of 
Africa, where mixed infections are not unknown, they might 
conceivably both occur in the same host, and here more especially 
since the geographical distributions coincide. 

T. pecaudi appears to have more claim to recognition in the field. 
Morphologically, the similarity between it and T. dimorphon is great, 
although it has been noted that the ‘ short * form of T. pecaudi maj' 
attain a greater breadth and carry a slightly better developed 
undulating membrane than is usual in the type strain, and both forms 
are present at the same time. Laveran, 3 however, states, ‘ these 
morphological differences do not suffice to differentiate the two 



340 


parasites/ It will be remembered that Dutton and Todd 6 were unable 
to adduce much information regarding the animal reactions in the 
course of the natural disease of the Gambia, except in horses. These 
correspond to what are manifested in 1 baleri ’ as outlined by 
Cazalbou. 7 In experimental animals the results coincide : the disease 
is acutely fatal in rats, guinea-pigs and dogs, though Laveran has 
noticed some slight variations in the mouse, and in all animals, as in 
dimorphon and congolense infections, splenic enlargement is common. 
One further point of difference might be cited ; in sheep and goats 
experimentally infected with T. pccaudi , the blood, though virulent 
on subinoculation, very rarely shows parasites; in T. dimorphon 
infection this is not usual. We have already referred to the variability 
of these animals under experimentation. 

The parasite which was obtained in the majority of cattle at 
Broken Hill corresponded to the original description given by Dutton 
and Todd of T . dimorphon , in so far that all three forms, including 
that with a free flagellum, were found, and the animal reactions were 
similar. Since the free flagellated and the small forms were not 
present simultaneously, it is improbable that we were dealing with 
T. pccaudi ; and as T. congolense does not appear to occur in a ‘ long * 
form it may also be negatived. 

3. Group including T. nanum , T. vivax and T. cazalboui. 

This group, constituted on the grounds of immunity enjoyed by 
the usual laboratory animals, presents difficulties in the way of sub¬ 
division, as from the very nature of the parasites they are more 
difficult of use in experimental observation, and hence less studied. 

Of the three species, T. nanum takes priority ; but, if we may 
judge from the very limited amount of work that has been possible, 
it is clearly separable from the other two on account of its morpho¬ 
logical features. Laveran 8 describes it as only measuring 10/1 to 
14/i in length, and this in conjunction with Balfour’s observations 9 
that two monkeys, two rabbits and a dog were not infected are held 
to substantiate the species. It is true that the morphological 
appearances strikingly recall the 'tadpole’ form of T. dimorphon ; 
but though the number of inoculated animals be small, infection, in 
some at least, should have resulted had this organism, to which 
laboratory animals are highly susceptible, been employed. Further 



34* 

Work may regain T. uanuni in the Sudan, when its position may he 
made more clear; until that time its specificity must remain 
questionable, though we incline to consider it as distinct from 
T. dimorphon„ and, therefore, to have no relation to the trypanosome 
of North Western Rhodesia which we have associated with that 
species* Further, its small dimensions preclude the possibility of its 
connection with either T. vivax or T . cazalboui . 

In July, 1906, Laveran 10 announced his belief that the trypanosome 
of 4 La Soumaya * was a new species, to which he gave the name 
T. cazalboui. Up to this time the impression had been created by 
various writers that the causal agent of La Soumaya was T . evansi , 
and, indeed, such statements had been made. Laveran and Mesnil 11 
say 4 ils ont la plus grande resemblance avec le trypanosome de la 
Mbori/ and they proceed to quote Cazalbou’s animal inoculations 
which resulted in the death of grey rats, mice and Sudanese dogs. In 
a later paper, published in May, 1907, Laveran 3 advances such 
additional evidence as to make the species incontestible. By 
morphology, animal reactions and cross-inoculations, T. cazalboui is 
clearly separated from the evansi and dimor phon groups, and from 
T. nantim , to which it is only related by the similarity of animal 
reactions. On all grounds it is a ‘good species/ and one which can 
be detected and classified in the field with comparatively little trouble. 

The same can hardly be said of T. vivax , as we yet know it, an 
organism described by Ziemann in 1905, a year before T . cazalboui 
was created, and given a specific name mainly on account of its 
rapidity of motion in cover-glass preparations. The morphology of 
this organism is in dispute : Ziemann 12 distinguishes it from T. brucei , 
though Laveran 13 was unable to note any difference from T. evansi 
in the film he examined ; and Schilling 14 contends that it is but a 
slightly more rapid form of T. brucei . This opinion is not reflected 
by Liihe, who writes 15 4 Ich selbst finde in einem mir iibersandten 
4 Originalpraparate die unter sich durchaus gleich gestalteten 
4 Trypanosomen kleiner wie Tryp . brucei .... mit nur schwach 
4 ausgebildeter undulierender Membran, wenig hervortrefenden 

*Wenyon (Report of the Wellcome Research Labs., Vol. 3) has apparently 
recovered T. nanum. We consider that one of the trypanosomes we have isolated 
from cattle, sheep and goats in North Eastern Rhodesia is closely allied to this 
species. (April 15th, 1909.) 



t Bandsaum derselben sehr kurzer oder vollig fehlender freier Geissel, 

‘ rundem Blepharoblast und niclit auffallig zugespitztem Hinterende.’ 

‘ Die Lange betragt nach Zic nann 18-26, bisweilen bis zu 30/*, die 
‘ Breite 2-2£/i ; der runde Blepharoblast liegt nahe dem meist etwas 
‘ zugespitzten Hinterende.’ To this we may add that Ziemann found 
difficulty in differentiating ‘ sexual * forms; an implication that 
this trypanosome is not subject to morphological variations of 
more than slight degree. Writing on Trypanosoma cazalboui^ 
Laveran says f its length, including flagellum, is 21//, breadth 1*5^. 

1 Nucleus oval and situated towards the middle. The centrosome 
‘ round and distinct is placed near the posterior extremity, which is 
‘ rounded and not pointed. Undulating membrane is very slightly 
‘ developed, being little folded as in T. lewisi 

A comparison of Liihe’s and Laveran’s descriptions of T . vivax 
and T . cazalboui , respectively, shows how little morphological 
difference there is between the organisms in stained film; and the 
similarity is accentuated when, referring to the movement in fresh 
preparation, Ziemann’s organism, named on account of its motility, is 
described as moving 1 “ wie einen Hecht ” in mehr oder weniger 
‘gerader Linie quer durch das Gesichtsfeld schiessen lasst;’ while 
T . cazalboui is said to be ‘ very active, moving sometimes on itself, 

* at others soon leaving the field like an arrow.’ 

The animal reactions of T. vivax have been imperfectly studied. 
Comparing the natural disease induced in cattle, sheep and goats, we 
can note no great difference from La Soumaya. Experimentally, 
Ziemann is quoted by Sander and Hennig 16 as having had the 
following results: 1 Graue Ratten, Tod nach 8-11 Tagen; Deutscher 
‘ Hund (?); einheimischer Schweine: nur leichte Erkrankung; 

‘ Esel: chronischer Verlauf . . . Ohne Erfolg: Katzen, Haus- 
‘ geflugeln, eine weisse Ratte.’ Nabarro in his analysis of Ziemann’s 
paper writes 17 : ‘ Dogs, cats and pigs were found not to suffer from 
‘ the natural infection.’ 4 A dog and a native sucking-pig developed 
f a slight temporary infection. White rats, geese, ducks, native hens, 
4 young turkeys, a native cat and an old pig were all refractory.’ 
Despite the limitations of Ziemann’s opportunities, it remains 
unquestionable that he was dealing with an organism showing marked 
lack of pathogenicity towards experimental animals. Laveran has 
not recorded the results following inoculation in grey rats, but 



343 

Cazalbou had previously noted their susceptibility to the trypano¬ 
some of La Soumaya. 

It is most certainly to be regretted that T. vivax has not been 
placed on a more substantial footing; but the evidence that is before 
us indicates strongly that it has few if any affinities with either the 
evansi or dimorphon group, and we are unable to see wherein 
T. cazalboui differs in any manner from what has been made known 
regarding T . vivax , and as further indicating the resemblance between 
these two, it may be added that Tabanidae are blamed by the German 
writer for transmitting his species in the Cameroons, and Cazalbou on 
more than one occasion lately has emphasised his belief that this 
family is concerned in the dissemination of La Soumaya in th'e Niger 
regions. 

Though they have never received specific designations, two other 
trypanosomiases of cattle deserve mention in connection with this 
group: that of Bruce, Nabarro and Greig at Entebbe, 18 and that 
described from Erythrea by Memmo, Martoglio and Adani. 19 In 
each instance the respective observers have noted the insusceptibility 
of laboratory animals to infection. The following paragraph sums 
up the morphology of the Erythrean organism :—‘ The trypanosome 
1 is morphologically like T. brucei or T. evansi , but is not more than 
‘ 24 fi long; free flagellum, which is fairly long, included. It is 
‘ extremely motile, like T. vivax of Ziemann. This trypanosomiasis 
‘ appears to be very virulent for ruminants, and thus differs from 
‘ typical surra. In many respects it resembles the disease described 
‘by Cazalbou in French Sudan under the name Souma. There is no 
tsetse in the infected area, and the suspected fly is a Tabanus or a 
* Hippobosca.' 

It appears to us that these two species are incapable of absolute 
diagnosis, save, perhaps, after a rigorous critical comparison. This 
to us in Africa is impossible, and we have consequently associated our 
second cattle trypanosome with the senior member. We would add 
that Broden 4 has also been struck by the unusual similarity between 
T. cazalboui and T. vivax , which he considers he has obtained in the 
Congo. 

Kambole, North Eastern Rhodesia, 

October 5///, igo8. 




344 


LITERATURE 


1. Montgomery and Kinghorn. Ann. Trop. Med. and Paras., 1908, 11, p. 97. 

2. Dutton and Todd. The Liverpool School Trop. Med., Memoir XXI, 1906. 

3. Laveran. Ann. Past., 1907, XXI, p. 347. 

4. Broden. Rapport sur la Lab. de Leopoldville, Bruxelles, 1906. 

5. Martin, Leboeuf and Roubaud. Bull. Soc. Path. Exot., 1908, 1, 6, p. 351. 

6. Dutton and Todd. Thompson Yates and Johnston Lab. Rep., 1903, Vol. 5. 

7. Cazalbou. Rev. Gen. Med. Vet., 1907, IX, p. 564. 

8. Laveran. C. R. Soc. Biol., 1905, p. 292. 

9. Balfour. Report of the Wellcome Research Labs., II, 1906. 

10. Laveran. C. R. Acad. Sc., July, 1906, p. 94. 

11. Laveran and Mesnil. Les Trypanosomes et Trypanosomiases, Paris, 1904. 

12. Ziemann. Quoted by 17. 

13. Laveran. Quoted by 15. 

14. Schilling. Quoted by 17. 

15. Luhe. In Mense, Handb. der. Tropenkrank, Bd. Ill, Leipzig, 1906. 

16. Sander and Hennig. Idem. 

17. Nabarro. Translation of Laveran and Mesnil, Trypanosomes and Trypano¬ 

somiases, London, 1907. 

18. Report of the Sleeping Sickness Commission, Nos. 4, 5 and 6. 

19. Quoted by 17. 



345 


EXPERIMENTS ON THE COMBINED 
ATOXYL - MERCURY TREATMENT 
IN MONKEYS INFECTED WITH 
TRYPANOSOMA GAMBIENSE 


BY 

ANTON BREINL, 

DIRECTOR OF TI1E RUNCORN RESEARCH LABORATORIES 

From the Runcorn Research Laboratories of the Liverpool School of 

Tropical Medicine 

(Received for publication 24 March y lgog) 

Prolonged experience of Sleeping Sickness treatment in man, and 
to a certain extent in experimental animals, has proved beyond 
doubt that Atoxyl by itself effects a really permanent cure only in 
comparatively few and exceptionally favourable cases of Sleeping 
Sickness in man, and if administered over a very prolonged period. 
Nearly all experiences with horses and cattle infected with the 
different strains of pathogenic trypanosomes lead necessarily to the 
conclusion that Atoxyl alone is insufficient for a successful issue of 
the treatment, but only prolongs life to a certain extent, and that 
finally the animal nearly always succumbs to the disease. 

Laveran’s 1 experiments showing that a combination of Arsenious 
Acid and Trypanred was superior on small animals to the use of 
either drug by itself, led the way for further advance in the treatment 
of Trypanosomiasis. Thomas and Breinl 2 after the introduction of 
Atoxyl recommended the combination of Atoxyl and Trypanred, the 
latter being then, after Arsenic, the best trypanocide known. 

The idea of a combined treatment in Trypanosomiasis was taken 
up later by Moore, Nierenstein and Todd, 3 and their extensive study 
of the experimental treatment of Trypanosomiasis was directed by 
the assumption that ‘ perhaps the recurrence in cases of trypanosome 
infection after treatment with Atoxyl might be due to some resistant 
stage of the parasite which survived the first treatment of Atoxyl, 



and later gave rise to recurrences of trypanosomes more or less 
refractory to Atoxyl treatment/ The Atoxyl was, therefore, 
followed by a second drug; the best results, experimentally, were 
obtained by using mercury salts, after the disappearance of the 
parasites from the peripheral circulation had been brought about by 
Atoxyl. In small animals, such as rats and rabbits, infected with 
T. brucei , the results were far superior to treatment by Atoxyl alone ; 
in large animals, as donkeys, on the other hand, the combined treat¬ 
ment with Atoxyl and Mercury was not found to be efficacious 
enough to be of practical value. 

Plimmer and Thomson 4 repeated the experiments on rats infected 
with Surra and Ngana, obtaining similar results to those of Moore 
Nierenstein and Todd, but using different mercury salts. In their 
hands the combination of Atoxyl and Succinamidc of Mercury gave 
the best results. In a further communication , 3 however, they state 
that, in small animals at any rate, Mercury has not given altogether 
satisfactory results, which fact they attribute to the small doses of 
Mercury. On enlarging the doses of Mercury, chronic kidney, and, 
to a lesser degree, liver lesions, were observed. They conclude, 
therefore, that more favourable‘results might be obtained in dealing 
with a more chronic trypanosome disease, such as Sleeping Sickness 
in man. 

Laveran’s and Thiroux’s 6 observations on the combined treat¬ 
ment of Atoxyl and Bichloride of Mercury on Surra in guinea-pigs 
are, although not very good, superior to the treatment by Atoxyl 
alone. 

According to Uhlenhuth, Hiibner and Woithe, 7, 8 the combined 
treatment on rats infected with Dourine was of considerable value. 
Criticising Laveran’s and Thiroux’s experiments, they remark that 
in guinea-pigs, Atoxyl treatment in general does not give good 
results, this being due to the fact that these animals very often die 
after one injection of Atoxyl without any apparent reason. 

The good results obtained by the combined treatment of Atoxyl 
and Mercury in experimental animals, justified its use in human 
Trypanosomiasis. The Segregate n Camps for the medical treat¬ 
ment of Sleeping Sickness in Uganda, gave this method a thorough 
trial . 9 

Mercury was administered in different forms and at different 



347 


intervals; in one series of cases concurrently with the Atoxyl, in 
another series some time after the last injection of Atoxyl. It is of 
interest to compare the different modes of administration and the 
results obtained either with Atoxyl alone or with Atoxyl followed by 
Mercury. The following methods have been used since the camps 
were started : - 


I. ATOXYL ONLY 

(a) 0’4 gm. every twentieth and twenty-first day. 

(b) 0*4 gm. every tenth and eleventh day. 

(V) 0'4 gm. increasing gradually every tenth and eleventh day 
up to 07 gm. 

(< d) Van Campenhout’s method (very similar to method ( c )). 

(< e ) 1 gm. every fifteenth and sixteenth day. 

(/) °‘6 gm. every fifteenth and sixteenth day. 


II. ATOXYL AND MERCURY 

Course of Aroxyl treatment lasting a month or six weeks, during 
which time at least 4 gm. of the drug were given, followed by— 

(g) Mercury perchloride, T l F grain, twice daily (Dr. J. Collyns). 

(h) Mercury perchloride, £ grain, hypodermically, for six doses 

spread over fourteen days (Dr. Collyns) 

(1) Metallic Mercury (Lambkin's cream), 5 minims once a week 
(Dr. C. J. Baker). 

(;) Intramuscular injections of 1 c.c. of a I per cent, solution 
of soluble mercury salts, repeated every five days 
(Drs. Goodliffe and Bayon). 

Combined simultaneous Atoxyl and Mercury treatment (Dr. van 
Someren)— 

(£) First day, Atoxyl, 1 gm.; Mercury perchloride, 0 01 gm. 
Second day, Atoxyl, 1 gm. On fourteenth day, Atoxyl, 
0*5 gm.; Mercury perchloride, o*oi gm. On fifteenth day, 
Atoxyl, 0*5 grn.,~ repeating every fourteenth and fifteenth 
day. 

(/) Same as above, except that one initial dose of 1 gm. Atoxyl 
is given, the remaining doses being 0*5 gm. 



348 


A comparison of the results obtained at the various camps with 
Atoxyl, and Atoxyl and Mercury treatment, during the period 
December, 1906, to November, 1907, is seen in the following table 
(Table VII, p. 8, of the report) : — 


Present State on February 29, 1908. 

Bus 
(Dr. Ci 

1 RO. 

allyns.) 

Atoxvl 

and 

Mercury. 
Method 
( g ) or (h) 

Kyagwk. 

(Dr. van Someren.) 

1 

Atoxyl and 
Mercury. 

1 Method (k) or (/). 

USOGA. 

(Dr. C. J. Baker.) 

Atoxyl I 
only. 
Method 
(ft) (C) (d) 

Atoxyl 
only. 
Method 
(ft), few (e) 

t 

Atoxvl 

ana 

Mercury. 

Method 

(0 


Per cent. 

Per cent. 

Per cent. 

Per cent. 

Percent. 

Improved 

8 

*9 

4°‘5 

35 

30 

Relapsed 

1 6 

x 5 

I I 

7 

5*5 

Continue in same state 

' 38 

62 

12*5 

3 

40 

Absent at time of examination 

1 3 

— 

21 

j 20 


Died . 

45 

1 

I 4 

*5 

35 

i 2 5‘5 

Number of cases 

382 

100 

328 

252 

1 

; 73 


The following table shows the comparative results from November- 
February, 1908, wherein the state of the disease is classified. 
A, meaning very early cases without symptoms, except gland enlarge¬ 
ment ; B, early cases with symptoms, itchy skin, &c.; C, advanced 
cases; D, very advanced cases (Table XVIII, p. 19, of the report).: — 





Ct .ass of Cask 

on Admission. 



Present State on 


A 

B | 

c 


D 

February 29, 1908. 




! 





Atoxyl 

Atoxvl 

Atoxvl 

Atoxjl 1 

Atoxyl 

Atoxvl 

Atoxyl 

Atoxvl 


only. 

j and | 

only. 

and 

only. 

ana 

only. 

and 


1 Mercury. 

Mercury. 

Mercury. 

Mercury. 


Per cent. 

Per cent. 1 

Per cent. 

Percent. 

Percent. 

Per cent. 

Percent 

. Percent. 

Improved 

4 2 

60-5 1 

35 

63 

1 24 

4 1 

7 

12 

Relapsed 

I 

3 i 

4 

4 

3 

7 

— 

— 

Continue in | 
same state t 

5 ° 

• 23 

46 

23 

56 

36 

53 

44 

Absent at time) 
of examination J 

3 

10 

5 

8 

5 

6 

8 

— 

Died ... 

4 

3‘5 

1 

10 

2 

12 

1 

IO 

32 

44 




.m 

I he method of treatment according to ‘ improved ’ cases is, 
therefore: — 

I. Method ( k) t 64 per cent. • 

II. Method (7), 58 per cent. 

III. Method (g), 39 per cent ' Atoxyl and Mercury. 

IV. Method (*), 34*5 per cent. ' 

V. Method (<b ), 34 per cent. ] 

VI. Method ( b\ 32 per cent. , Atoxyl alone. 

VII. Method (/), 10 per cent. J 

Bohne 10 describes one case of trypanosome fever which improved 
markedly under the combined Atoxyl-Mercury treatment, but the 
time of observation is too short to conclude that this patient has been 
definitely cured. 

Broden and Rodhain, 11 on the other hand, come to the conclusion 
that the combined treatment did not prove superior at all to a 
treatment with Atoxyl alone. 

As Moore's, Nierenstein's and Todd's work was mostly carried out 
on animals infected with T. brucei , it seemed advisable to repeat the 
work on monkeys infected with T. gambiense. Six animals 
(Cercopithecus callithricus) were inoculated with our laboratory 
strain of T. gambiense , and after varying time subjected to treatment. 
Acetylated Atoxyl, which, in former experiments, had proved itself 
less toxic than Atoxyl, was used. 

Experiment I.— Cercofithecus callithricus , 2 , weight 2 k. 700 gm., was 

inoculated on April 28th with T. gambiense. At the time of the third relapse, on 
June 1 st, treatment was begun after the animal had shown typical signs of the 
infection ; the animal was markedly anaemic, and had lost over 300 gm. in weight. 
On June 1st and and, an injection of o*i gm. of acetylated Atoxyl was given. The 
parasites disappeared after the first injection. On June 10th and 18th, the Atoxyl 
was followed by an intra-muscular injection of 0*05 gm. of Sublimate. On July and 
and 3rd, the Atoxyl injection, and on July 15th and 16th the injections of 
Sublimate were repeated. Up to November, daily examinations of the blood were 
made, and since then the peripheral blood has been examined twice weekly. No 
trypanosomes have been seen since June 1st, the day of the injection. The blood 
count is normal (March 7th). The animal has increased in weight to a k. 845 gm., 
which has been maintained with slight variations since the middle of November. 

Experiment II.— Cercofithecus callithricus , 2 , weight a k. 700 gm., was 
inoculated on April 28th with T. gambiense. The animal never showed parasites 
in great number. On June 16th, treatment was begun, as the symptoms of the 
disease became more and more marked. On June 6th and 10th, 0*1 gm. of 
acetylated Atoxyl was administered; these injections were followed on June 18th 
and 19th by 0*05 gm. of Sublimate, intra-muscularly, in the gluteal region. The 
second injection of Sublimate was followed by complete paralysis of the hind leg, 

AA 



which was most probably due to an injury of the sciatic nerve through the injection 
This paralysis, however, passed off very gradually, and after two and a half months 
the mobility of the leg was normal. On July 2nd and 3rd, the acetylated Atoxyl 
injections were repeated, and on July 15th and 16th the intra-muscular injections 
of Sublimate. The animal is still alive, and parasites have never been seen since 
the first inoculation. The blood is normal, and the weight is at present 
ak. 895 gm. 

Experiment III .—Cercofithecus callithricus , 9 , weight 3 k. 600 gm., was 
inoculated on April 28th, 1908, with T. gambiense. Treatment was started on 
June 6th, the time of the third relapse. 0*15 gm. of acetylated Atoxyl was 
administered. This injection was repeated on June 10th, and followed on 
June 18th and 19th by intra-muscular injections of 0*075 8 m * Sublimate. The 
Atoxyl injections were repeated on July 2nd and 3rd, and the Sublimate injections 
on July 15th and 16th. This animal is still alive; parasites have never been seen 
since the treatment was started. The blood count is at present normal, and the 
weight, which had increased by July 21st to 3 k. 850 gm., has remained constant 
since November at 3 k. 600 gm. 

Experiment IV .—Cercofithecus callithricus, 3 , weight 1 k. 800 gm., was 
inoculated on April 28th. Treatment was started on June 3rd with injections of 
o-igm. of acetylated Atoxyl, followed by an administration on June 8th of 
o*oi gm. of Sublimate in pill form, by the mouth. A very severe attack of diarrhoea, 
which lasted from June 9th—12th, followed this medication ; the stools were slimy 
and haemorrhagic. On June 18th and 19th, o*i gm. of acetylated Atoxyl was given, 
followed on June 23rd by one injection of 0*05 gm. of Sublimate. On July 2nd 
and 3rd, o*i gm. of acetylated Atoxyl was given, followed on July 15th and 16th 
by o*oi of Sublimate, intra-muscularly. The Sublimate injections were not followed 
by any untoward effects. The animal is still alive ; parasites disappeared after the 
first injection of Atoxyl, and have never been seen since. The blood is normal, 
and the weight 2 k. 100 gm. 

Experiment V .—Cercofithecus callithricus , 9 , weight 1 k. qoo gm., was 
inoculated on April 28th. At the time of the second relapse on June 3rd, treatment 
with acetylated Atoxyl was begun. Two injections of o*i gm. on June 3rd and 4th 
were followed by an administration of o*i gm. of Sublimate in pill form by the 
mouth. In this case, as in Experiment IV, a very severe diarrhoea with slimy and 
haemorrhagic stools occurred, which, however, passed off in a few days. The 
Atoxyl injections were repeated on June 18th and 19th, followed on June 26th and 
27th by an intra-muscular injection of 1 c.c. of Donovan’s solution. On July 2nd 
and 3rd the Atoxyl injections were repeated, followed on July 15th and 16th by 
an injection of 1 c.c. of Donovan’s solution. The parasites disappeared from the 
peripheral circulation of the animal after the first injection, and were never seen 
again. The animal was found dead on July 17th. The post-mortem showed fatty 
degeneration of the heart ; the spleen was slightly enlarged, of firm consistence, 
with well-marked hypertrophic malpighian follicles. The kidneys were enlarged 
and markedly congested, the cortical substance was not defined from the medullary 
substance, and haemorrhagic stripes intersected the yellow cortical substance. 
Microscopically, the liver showed typical fatty degeneration of the liver cells; the 
kidneys showed on section a well-marked parenchymatous nephritis. The death 
of the animal was due in all probability to Mercury poisoning. 

Experiment VI .—Cercofithecus callithricus , 9 , weight 2 k. 750 gm., was 
inoculated on June 6th with T. gambiense. Treatment was started on June 18th 
with a view to ascertaining whether a short treatment at an early stage can effect 
a permanent cure. o*i gm. of acetylated Atoxyl was given, and the same dose 
repeated on June 18th. The parasites, which were fairly numerous before treat¬ 
ment, disappeared promptly. In this case, however, the acetylated Atoxyl 



produced a harmful effect. The doses which proved perfectly harmless in the 
foregoing experiments caused the whole complex of symptoms of a typical Atoxyl 
poisoning. Tremors occurred all over the body, and, at the same time, a severe 
slimy diarrhoea started. These symptoms, however, passed off in the course of a 
week. On June 26th and 27th, injections of 1 c.c. of Donovan’s solution was given. 
This animal is still alive, weighing at present 3 k. 100 gm. The blood is normal; 
parasites have never been seen since treatment was begun. 


The foregoing experiments show that in five cases out of six, 
the administration of Acetylated Atoxyl and Sublimate, and 
Donovan’s solution, in monkeys ( Cercopithecus callithricus), has 
effected a complete cure. 

If we consider that in our experiments Cercopithecus callithricus 
usually succumb to an infection of T. gambiense in the course of 
40-60 days, the results of Atoxyl-Mercury treatment in monkeys 
infected with T. gambiense must be looked upon as very encouraging 
indeed. 

The length of time which has elapsed since treatment was 
discontinued is probably sufficient to permit of the animals being 
considered as definitely cured of the disease. 

Concerning the action of the combined treatment by Atoxyl and 
Mercury, a conclusive opinion can hardly yet be expressed. Mercury 
has proved beyond doubt to have not the least effect on trypanosomes 
in the peripheral circulation. It is, therefore, very tempting to accept 
Moore’s, Nierenstein’s and Todd’s suggestion of its action on a 
secondary resistant form of the parasite which it destroys; but 
whether we accept this hypothesis or suppose that the Mercury acts 
merely as an internal disinfectant after the destruction of the 
parasites by Atoxyl, the success of the combined treatment seems to 
depend upon the administration of the two drugs either concurrently 
or in rapid succession. 



LITERATURE 


1. Laveran. Traitement mixte des Trypanosomiases par l’acide ars^nieux et le 

trypanroth des infections dues aux Trypanosoma gambiense. Compt. rend, 
de l’Acad. d. Scien. 1905, T. 140, p. 287. 

2. Thomas and Breinl. Trypanosomes, Trypanosomiasis and Sleeping Sickness. 

Memoir XVI, Liverpool School of Tropical Medicine, 1905. 

3. Moore, Nierenstein and Todd. A note on the therapeutics of Trypano¬ 

somiasis. Annals of Tropical Medicine and Parasitology, Vol. I, No. 1, 1907. 
Concerning the Treatment of Experimental Trypanosomiasis. Annals of 
Tropical Medicine and Parasitology, Part I, Vol. I, No. 2, p. 275, 1907, and 
Part II, Vol. II, No. 4, 1908. 

On the Treatment of Trypanosomiasis by Atoxyl, followed by a Mercury salt, 
&c. The Bio-chemical Journal II, No. 5, 6, 1907. 

4. Plimmer and Thomson. A Preliminary Summary of the results of the 

Experimental Treatment of Trypanosomiasis in rats. Proceedings of the 
Royal Society, Series B, Vol. 79, 1907. 

5. Plimmer and Thomson. Further results of the Experimental Treatment of 

Trypanosomiasis in rats. Proceedings of the Royal Society, Series B, 
Vol. 80, pp. 1-11, 1908. 

6. Laveran et Thiroux. Recherches sur le traitement des Trypanosomiases. 

Annales de l’lnst. Pasteur, p. 97, 1908. 

7. Uhlenhuth, Hubner und Woithe. Experimentelle Untersuchungen iiber 

Dourine mit besonderer Beriicksichtigung der Atoxylbehandlung. Arb. a. d. 
Kaisl. Gesundheitsamte, Bd. XXVII, 1907. 

8. Uhlenhuth und Woithe. Nachtrag und Schlussbericht. Arb. a. d. Kaisl. 

Gesundheitsamte, Bd. XXIX, Heft. 2, 1908. 

9. Gray. Quarterly Report on the Progress of Segregation Camps and Medical 

Treatment of Sleeping Sickness in Uganda. Sleeping Sickness Bureau, 
London, 1908. 

10. Bohne. Ein Fall von Trypanosomenfieber mit langer Dauer und seine 

Behandlung. Verhandlungen der deutschen Tropenmedizinischen Gesell- 
schaft. Arch. f. Schiffs und Tropenhygiene, Band XII, Beiheft 5, 1908. 

11. Broden et Rodhain. Rapports sur les travaux etc k Leopoldville, 1907- 

1908. Hayez, Bruxelles, 1908. 



353 


DRUGS FROM THE CONGO 

BY 

PROSPER H. MARSDEN, F.C.S., 

LECTURER IN MATERIA MKDICA AND PHARMACY IN THE UNIVERSITY OF LIVERPOOL 


(Received for publication 25 March , 1909J 

In a former article* I have described some of the native African 
Drugs in the Museum of the University of Liverpool and spoken of 
the importance of a closer study of the remedies which have been in 
use among the natives, perhaps for centuries, for the treatment of 
disease. 


MOUINDU OR MUINDU 

On July 19th, 1906, I received from Mr. Robert Newstead a 
packet of drugs which were brought to this country by Prof. J. L. 
Todd, and had been sent, together with Palabanda and other drugs 
from the district of Banana, to the late Dr. J. Everett Dutton, on the 
17th February, 1904, by the Commandant Du Camp of the lower 
Congo, in response to enquiries by Dr. Dutton concerning native 
remedies against Sleeping Sickness. A further sample of the same 
drug was given to me in March, 1909, by Dr. J. W. W. Stephens, 
which had been sent by Dr. E. Etienne from the same district. 

Macroscopic Characters of the Drug . 

The material consisted of a piece of stem 22 cm. long by 3 cm. in 
diameter, together with leaf stalks and detached leaves, of the 
appearance of which fig. 1 gives a good idea. 

The cortical portion which was readily removed from the hard 
woody portion had an astringent taste, followed by a persistent 
bitterness. No part of the drug had any characteristic odour. The 
outer layer of the cortical portion was velvety to the touch, 
longitudinally furrowed, with occasional transverse cracks, and 


- Prosper H. Marsden : Notes on the Drugs of West Africa. Quarterly Journal of 
the Liverpool University Institute of Commercial Research in the Tropics, Vol. Ill, No. 6. 




ww 




numerous transverse fenticels This velvety layer could by easily 
removed in - the linger nail, showing £ dark red brown hard .hark 
beoeaiJi. 

A .transverse section oi the stem showed he Outer bar); 10 be 
IS trail, thick and the inner sed-bfowii by a 3 to 4 min. thick; in 
this V-sfcfcyJ mahid thb mnci 

side. .T in’ bark had genenvOy ' V- .yhoji .with ftmy VHbjrcs 

ohO'ftdiib hr the site -of"ittigiurc.' '7. '. ' 


1- ■. ■i^Tvkttivts. ui«i btem -of Mumdit 


Ibe wood, ry rra at the thickest part, was hard, yel!o\vish-\vKit* 
n-oh five. rings and minute pith, and with a,powerful.fens the lacunae 
■ <0,, 'd !>e meri ".Inch are' rhartacteristic of the gemts..S7r>r//w.t 







J 55 

As the bark of Muindu has some resemblance to that of the false 
Angostura Bark ( Strychnos Nux Vomica , Lin.) described about half 
a century ago,t and both Muindu and Palabanda have something in 
common with Sassy Bark ( Erythrophloeum guineense , Don.), it was 
thought that in view of the medicinal use of these an examination 
of Muindu might be of value. 

The sample from Dr. Etienne consisted of three pieces of branches 
of the plant, which he described as a shrub growing in the plains. 
The largest of the branches was some 50 cm. in length by 2'5 cm. in 
diameter at the lower end, which had the velvety appearance of the 
older stem above mentioned. The younger branches were smoother, 
and had sharp thorns 5 mm. to 2 cm. long irregularly distributed 
upon them. There were no leaves with this specimen, but the 
thinnest branch had evidence of leaf scars, and was of the same 
colour and appearance as the leaf stalk of the Muindu received from 
Dr. Todd. A note by Dr. Etienne states that the natives scrape off 
the liber and macerate in treaty rum. Of this preparation a few 
drops are instilled into the eye for Sleeping Sickness. 

A tincture was prepared from the bark by maceration in 60% 
alcohol of a strength of one drug in ten of menstruum. This was 
tested on frogs by Dr. C. O. Jones in the Department of Bio- 
Chemistry of the University with negative results. 

This tincture was shaken out with ammoniated chloroform three 
times, and the chloroformic layer being allowed to evaporate to 
dryness gave a residue which upon treatment with sulphuric acid and 
potassium bichromate failed to give the usual reaction for strychnine. 

Microscopic characters of the Bark of Muindu. 

A transverse section of the bark showed that the velvety layer 
had an indefinite appearance. 

The suber is formed of flattened thin-walled cells, the cortical 
parenchyme consists of polygonal cells having thin walls and inter¬ 
cellular spaces, and containing a brown material which gave the 
reaction for tannin with ferric chloride. 

Although, as above mentioned, I was unable to obtain the reaction 
for strychnine from the tincture of Muindu, upon flooding a transverse 


fPlanchon et Collin. Le* drogues simples d'origine v^getale, Vol. T. p. 662. 





section of the bark with siiljJho-vaaadt'; acid the customary -violet 
coloration -iue to strychnine was afforded.. This was not given by a 


.are f^und a; bp-groups bi ; s^erc'i'jchyoiie 

w;Ui small luiuem These groups arc more numerous towards the 
toner Side, when-iiiey avaunt? a tfuriclrbiigiilat shape nod.are. arranged 
very dosfcly u.gejhv'jr (%. r>. <u longthiduial section these setak-ren- 
chvoiafcoust'fibres .cr seen to he- elongated longitiidviialK- fig. $>. 


I'li*.. j -i -f.t ifOH f'f hpyK r,f ’vhsuKUi. 


Cftetxiiti'y'tvf t.fa Uiifrdi ,tiht#4ir 

rhe leaves have not the character of those of the genus StrycJtu.**, 
but as they were detached when received, j» is possible..that - they- are' 
not from the same plant *5. the 'Stem and branches. A photo¬ 
micrograph of the mid-rib of .one of the leaves is seen m fig. but m 
view of the statement of Dr. Ttietme' that the bark oi the:plant >ya.s 
the pari used i'n 'medicine, the leave- -.'. ere not further jnvesligatvH 





Krr*, 3.—Longitudinal iteU0f> or bark of AJiumiu. 


rtewM 


F?g. Tvaris^ei se section of mid-rih of leaf 
of Mumdu. 





A GREGARINE PARASITIC IN THE 
DOG-FLEA, CTENOCEPHALUS 
SERRATICEPS 


BY 

E. H. ROSS, M.R.C.S. England, L.R.C.P. London, 

HEALTH OFFICER, SUEZ CANAL, EGYPTIAN PUBLIC HEALTH DEPARTMENT 


(Received for publication 29 March , igog) 

The excuse for describing this parasite which inhabits the 
alimentary tract of the dog-flea must be in the fact that much work is 
now being done with fleas generally, and some confusion may be saved 
to others, working at these insects, if some details of its various phases 
are related. 

This parasite was found, in varying numbers, in the fleas of two 
fox-terrier dogs, mother and son, which had lived in Port Said nearly 
all their lives. Its stages follow the well-known gregarine type 
frequently found in Nematodes and Culicids. It is a Cephalin showing 
a well-marked cycle of sporogony, all the stages of which are 
completed within the body of the flea, its host; while, like many 
other gregarines, the cycle of schizogony, so far as can be found out, 
is wanting. 

It is convenient, for the sake of description, to begin with that 
phase of the cycle which is first seen in the flea. 

I. THE EARLY TROPHOZOITE 

This stage, as in similar Gregarinidae found in the Worms, 
Echinoderms, and Ascidians, is the first resting phase of the parasite 
in the flea. It is probably caused by the direct infection of a stomach 
cell by a sporozoite derived from the rupture of the sporocyst, the 
sporozoite having been eaten by the flea larva when crawling on the 
dogs back. But it is possible that it may be derived from the 
merozoite of a cycle of schizogony which has taken place either 
outside the host or within the stomach of the flea-larva ; up to the 
present time, however, no traces of such cycle have been noted. 



The early trophozoite, then, is a small circular cell embedded 
between the pyriform epithelial cells lining the stomach of the flea. 
As many as twenty-five may be found in one flea situated near the 
proventriculus, but occasionally near the pylorus. They arc 
frequently found in pairs. These cells contain large refractile 
granules, and a readily staining nucleus. The whole cell stains before 
the stomach cells when the organ containing it is placed upon agar 
having an aniline dye in suspension. In clear specimens a slender 
process can be detected, by which the parasite is attached to the 
remains, probably, of its trophic cell that has been destroyed by it. 
The relation of this early form to the rostrated trophozoite next to be 
described is not merely conjectural, because it is only found in young 
infected fleas and in those containing older trophozoites which cannot 
be mistaken. Besides, the granules in all the early stages are very 
characteristic. The fact that these early forms are only found in 
young flea imagines makes it highly probable that this parasite is 
ingested by the larva, for in older fleas the more highly developed 
phases of sporogony only are to be found, and it is unusual to see 
parasites in other than contiguous stages in the same flea. The age 
of a flea may be roughly estimated by the degree of growth of the 
ova in the ovisacs in the females, and by the degree of development 
of the spermatozoa in the vesiculae seminales in the males. In the 
dog-flea the absence of spermatozoa in the spermatheca of the female 
is almost certain evidence of the extreme youth of the imago, 
because fecundation by the male takes place very soon after the 
metamorphosis of the female is completed; and in the male the 
spermatozoa within the vesiculae seminales are tied together by their 
heads into compact bundles very soon after the imago has hatched 
from the nymph. 

II. THE TROPHOZOITE AND ITS DEVELOPMENT 

During this phase the parasite develops a well-marked epimerite. 
In its growth it may reach a size equal to ten times that of a stomach 
epithelial cell. It is pear-shaped, and is fixed by its epimerite to the 
lining membrane of the stomach. The body of the cell is divided, in 
the early stages, by a horizontal septum into two nearly equal halves, 
the protomerite and the deutomerite. As the cell grows it becomes 



more highly granular and consequently darker to transmitted light, 
and the septum less distinct. It also begins to lose its pyriform shape, 
and slowly reverts to its circular form. When full grown it is circular 
and the horizontal septum has vanished, while it is full of large 
refractile granules and the epimerite has disappeared. These 
trophozoites are usually found in pairs within the stomach and 
adhering to its wall. 

III. THE FORMATION OF THE SPORONT 

The epimerite or rostrum has completely disappeared, and the 
parasite is circular. The next phase seen of this particular parasite 
is that of the association and encystment of two sporonts. The cyst 
is embedded in the stomach wall, and consists of a thick fibrous 
capsule. The gametes appear as small granules. The analogy of 
similar sporonts parasitic in other members of the Arthropoda shows 
that they are formed in the following manner:—A male and female 
sporont conjugate and become encysted in their mother tissue A 
nuclear spindle is formed from a small portion of the nucleus of the 
sporont and divides, the remainder of the nucleus degenerating. The 
spindle then produces daughter nuclei by mitosis and they again 
divide, until a number of nuclei are formed which bud off the surface 
of the sporont. Each budded nucleus is then the primary sporoblast, 
and ultimately becomes the male or female gamete according to the 
sporont from which it was originally derived * Whether in this 
instance, the male gamete becomes flagellated or not, it is impossible 
to say, but in the specimens examined in this stage no flagella were 
ever seen; however, the cephalonts are very difficult to stain, as they 
are surrounded with such a thick fibrous wall. The males are said to 
burst their way into the female half of the cephalont, and there fertilise 
the female gametes. 

The cephalont is a very remarkable looking cell embedded in the 
stomach wall. It is large, frequently thirty times the size of a stomach 
epithelial cell, and its two granular, male and female, halves make it 
very characteristic and distinct. It can be rolled when the stomach is 
pressed under the cover-slip, but I have not succeeded in staining the 
gametes, as sufficient force to rupture the cephalont causes their 
destruction. 

* I am indebted to Professor Minchin's article on the Sporozoa, in Lankesters 
Zoology for the description of this process. 



IV. THE FORMATION OF THE OOCYST 


After the fertilisation of the female gametes the male half of the 
cephalont degenerates, and the female portion grows until it fills the 
whole cell, which then separates from its attachment to the stomach 
wall and becomes free in the cavity. Each female gamete grows 
in size after fertilisation, while the fibrous cell-wall thins with the 
distension. The gametes, or zygotes as they must now be called, are 
highly refractile bodies, each about the size of a red-blood corpuscle. 
They are packed tightly in the fibrous capsule or sporocyst, which 
resembles a pomegranate that has been cut through the middle with 
a sharp knife. It is about fifty times the size of a stomach cell, and 
if the stomach wall be ruptured it may be extruded, transferred to a 
clean slide, burst, stained and examined. But under natural 
conditions it passes down to the pylorus, through which it is too large 
to move, and it ruptures, and the contained zygotes escape into the 
intestines (Malpighian tubes) under the influence of the peristaltic 
action. 


V. THE DEVELOPMENT OF THE SPOROBLASTS 

If a cyst be expressed from the stomach, crushed on a slide, and 
the sporoblasts stained by any method giving the Romanowski result, 
they will be found to be lanceolate in shape but of somewhat irregular 
contour. Each one has some chromatin, but this varies in amount 
from a minute dot to an extensive streak. The cell-wall is glistening, 
and appears almost chitinous, while there is some very feebly staining 
cytoplasm. But they have a high osmotic index, for if the intestine 
containing them be placed unruptured upon agar spread on a glass 
slide and containing polychrome methylene blue, according to the 
method described in the Journal of Physiology, for September 16th, 
1908, by H. C. Ross, it will be noticed that the living sporoblasts 
accept the stain before the cells forming the lining to the tube. But 
even these cells accept the stain more readily than the hepatic cells, 
or the epithelial cells of the stomach or of the salivary glands. They 
are, therefore, very susceptible to external influences. 

After the cyst has ruptured the sporoblasts pass into the 
intestine, where they undergo a still further change. They become 
barrel-shaped sporocysts; the contour is regular, and in the fresh 



state they are yellow and roll up and down the Malpighian tube with 
the flow of its contents. Sometimes they are very numerous, and can 
be seen at once on examining the intestines. 

VI. THE FORMATION OF THE SPOROZOITES 

While in the intestine the barrel-shaped sporocyst gives rise, in its 
interior, to eight rod-shaped sporozoites. These are at first found tied 
together by their ends like a bundle of cigars or bananas, so that when 
viewed from above the sporozoite gives the appearance of eight small 
separate circles within a circle. Up to the present it has been 
impossible to separate these sporozoites or to stain them, but when the 
sporocyst passes into the rectum of the flea it ruptures, and the 
sporozoites are set free with the faeces. The ultimate destination of 
the sporozoites has not been traced, but they are passed with the 
faeces. Whether, as stated before, there is a further cycle of 
schizogony, or whether the sporozoites are eaten directly by the flea 
larva and a new cycle of sporogony started, it is not possible to say. 
But the flea larva is difficult to obtain, and still more difficult to 
dissect. During two years’ work with dog fleas the number found 
infected with this parasite was 38 per cent. 

I suggest as a name for this Gregarine, Gregarina ctenocephali 


cams. 




365 


THE ACTION OF ARYL-STIBINIC ACIDS 
IN EXPERIMENTAL TRYPANOSOMIASIS 


BY 

ANTON BREINL, 

DIRECTOR OF THE RUNCORN RESEARCH LABORATORIES OF THE 
LIVERPOOL SCHOOL OF TROPICAL MEDICINE 
AND 

M. NIERENSTEIN, 

J. W. GARRETT INTERNATIONAL FELLOW, AND RESEARCH DEMONSTRATOR, LIVERPOOL 
SCHOOL OF TROPICAL MEDICINE 

Front the Runcorn Research Laboratories of the Liverpool School of 

Tropical Medicine 

(Received for publication 3 April , igogj 

Plimmer’s and Thomson's important discovery of the trypanocidal 
action of Antimony, an element chemically closely allied to Arsenic, 
was the starting point for extensive investigations on the value of 
different Antimony compounds in the treatment of Trypanosomiasis. 
In their first experiments 1 they used Potassium antimonyl tartrate, 
and observed that this drug destroyed the trypanosomes in the 
peripheral circulation more rapidly than Atoxyl. The injections 
caused neither pain nor inflammatory changes of the tissue. In their 
experiments, out of twenty-five rats only four showed recurrences; 
nine lived for over two hundred days, and nine considerably over one 
hundred days; the remaining three died without any symptoms of 
Trypanosomiasis, and in none were trypanosomes found after death. 2 

Mesnil and Brimont 3 were able to confirm Plimmer's and 
Thomson's observations concerning the powerful trypanocidal action 
of Potassium antimonyl tartrate. Their experiments on several 
strains of pathogenic trypanosomes, however, proved that after one 
injection the parasites usually reappeared, sometimes after a very 
short time. In Ngana infected rats, as many as nine relapses were 
observed, and the drug had always the same transitory effect; most of 
the animals died finally either with or without parasites after 
discontinuation of the treatment. Their results in animals infected 
with T. roansi y were more satisfactory; and it is specially noteworthy 

BB 



3 66 


that an Atoxyl-resistant Surra strain reacted to Antimony in the 
same way as a normal strain. A preventative action was only noticed 
if the drug and the parasites were injected simultaneously in twc 
different distant places. 

In our hands, 4 Sodium antimonyl tartrate did not give such 
promising results in the treatment of rats, a fact which might be due 
to the use of an especially virulent strain of T. equiperdum. One 
horse injected with a strain of cattle trypanosomes brought back from 
the Congo, and one donkey infected with an Atoxyl-resistant strain 
of Ngana, were treated with Sodium antimonyl tartrate in fairly large 
doses. In both cases the drug caused the prompt disappearance of 
the parasites; the interval between the relapses, however, became 
shorter and shorter after each injection, and finally both animals 
succumbed to the disease. 

Manson 5 was the first to administer Sodium antimonyl tartrate to 
a case of Sleeping Sickness. As Atoxyl given in large doses did not 
seem to have any effect on the trypanosomes, a treatment of Antimony 
in small doses was begun. It caused the disappearance of the 
parasites, but eighteen days afterwards parasites were again seen in 
the peripheral circulation. As the injection had caused intense 
irritation and pain, two grains of Antimony were given by the mouth ; 
this was followed by nausea, and seemed to increase the mental 
depression of the patient. Antimony treatment was discontinued, 
and Atoxyl again given. 

For further experiments Mesnil and Brimont 6 used mice infected 
with different laboratory strains of pathogenic trypanosomes. 
According to their results they were able to separate the strains into 
two groups. The parasites of the first group disappeared after a 
single injection: Surra and Dourine belonging to this group. In the 
second group, to which all other strains belong, the parasites 
disappeared after the injection of Tartar emetic, but only to reappear 
within a few days. The negative phases after each injection became 
shorter and shorter, and the animals finally succumbed to the disease. 

In the discussion, Laveran 7 *• 6 states that in his hands, Tartar 
emetic did not prove very satisfactory in guinea-pigs, as after the 
rapid disappearance of the parasites following the first injection, 
relapses occurred very frequently. Sulphide of Antimony, in his 
experience, is much less active than Sulphide of Arsenic (orpiment). 



367 


Uhlenhuth and Woithe 8 repeated the experiments with Sodium 
antimonyl tartrate on twenty-seven rats infected with T. equiperdum , 
but their results were as discouraging as those with Arsenious acid. 
Even repeated injections of 0*003—0*005 gramme of the Sodium salt 
and 0*002—0*003 gramme of. the Potassium salt were unable to effect 
even temporary disappearance of the parasites from the blood. 

Broden and Rodhain 9 used soluble as well as insoluble compounds 
of Antimony in Sleeping Sickness treatment The hypodermic 
injections caused great irritation and pain, and were followed, even 
when given intramuscularly, by large swellings; this reaction 
persisting for some days, and only disappearing after one .week. 
Therefore, the drug was administered intravenously. A dose of 
0*07 gramme was sufficient in some cases to cause the parasites to 
disappear, but they very soon reappeared in the peripheral 
circulation. These observers recommend a dose of 0*1 gramme of 
Tartar emetic. This dose given intravenously did not usually cause 
any severe symptoms; it was followed sometimes by profuse 
perspiration and vomiting. After repeated injections the patients 
usually lost their appetite and complained of general malaise. On 
interruption of the treatment these symptoms passed off. They were 
able to confirm the rapidity of the destructive action of Antimony on 
the parasites, but remark that, 1 Ces constatations doivent nous imposer 
une extreme reserve dans l’appreciation de la valeur de Tantimonie 
dans le traitement de la trypanosomiasis humaine et exigeront une 
experimentation longue et patiente/ 

They, however, place, for the present at least, the soluble Antimony 
compounds on the same level as Atoxyl. 

Broden and Rodhain, as well as Martin and Darre, 10 combined the 
Atoxyl treatment of Sleeping Sickness patients with injections of 
soluble Antimony compounds with very encouraging results. 

Good results of Antimony treatment in experimental Syphilis are 
recorded by P. Salmon, 11 and Broden and Rodhain 9 were able to 
confirm the beneficial effect of Antimony in human Syphilis. 

Plimmers and Thomson’s observations on the rapid action of 
Potassium antimonyl tartrate on trypanosomes in the blood of infected 
animals seemed to justify an attempt to prepare an organic Antimony 
compound analagous to Atoxyl. Moreover, a comparison of the 
effects of injections of Sodium arsenate with those of Atoxyl made it 



368 

probable that injections of the Sodium salt of Amino-phenyl-stibinic 
acid would be far less irritating and permit the introduction of a larger 
amount of Antimony into the organism without toxic symptoms. 

After many unsuccessful attempts, we succeeded in preparing the 
p.m. and o. amino-phenyl-stibinic acids. *The action of the p . and m. 
compounds has been studied on experimental animals infected with 
various laboratory strains of pathogenic trypanosomes; the o. 
compound was, after a few tentative experiments, given up as 
impracticable. 


I. EXPERIMENTS WITH m. AMINO-PHENYL-STIBINIC ACID 

m. amino-phenyl-stibinic acid was used in the form of its acetylated 
derivative. This latter proved itself in the first experiments the less 
toxic, and produced no appreciable irritation at the site of inoculation, 
whereas the m. compound caused marked swellings and abscesses. 

A. Rats. 

Medium-sized rats of the weight of 180-220 grammes were used 
for the following experiments. Untreated animals succumbed to an 
infection of T. brucei in 4-6 days on the average. Treatment was 
usually started when numerous parasites were present in the peripheral 
blood. In a few experiments the compound was injected only at a 
late stage, some hours before death; but then the animal always died 
from the infection. The toxic dose was found to be 075 c.c. of a 
5% solution ; an injection of 0*5 c.c. of the same solution corresponding 
to 0*025 gramme of the drug was usually well borne. No abscesses 
were noticed at the site of injection. After the administration of this 
dose the parasites disappeared from the blood in 12-16 hours. Smaller 
doses were only able to effect a disappearance of the parasites from 
the blood, when repeated. The parasites, however, often reappeared 
after a comparatively short time ; a further injection of the drug again 
resulting in the disappearance of the parasites. 

Table I shows the details of the result of treatment on thirty-three 
rats*, only one rat is still living after 136 days. 









37° 


B. Dogs. 

Two dogs were used for the experiments (see Table II). In 
experiment 245 the dog was treated with Sodium-amino-phenyT 
stibinic acid ; in experiment 244 with the acetylated derivative of the 
same compound. In the former experiment severe abscesses resulted 
from the injections, and after a very short time the animal succumbed 
to the toxic effects of the drug. The post-mortem examination 
revealed a haemorrhagic nephritis. 

The subcutaneous injections of the acetylated compound, on the 
other hand, did not cause any irritation. Only frequently repeated 
large doses effected a disappearance of the parasites from the blood. 
Very soon, however, trypanosomes reappeared again. The animal 
succumbed to a severe toxic haemorrhagic nephritis due to the 
Antimony. 

C. Rabbits. 

Six rabbits were inoculated with T. brucei , and after the disease 
had become well established treatment was begun. But even 
prolonged administration of fairly large doses—o* 1 gramme per 
injection—was not able to cope with the disease, and all the animals 
succumbed to the infection. Although parasites were very rarely 
seen in the peripheral blood, the well-known symptoms of a Ngana 
infection were more or less pronounced during the whole course of the 
treatment. 

D. Guinea-pigs. 

Treatment was only attempted in the case of three guinea-pigs, as 
it was soon apparent that these animals did not bear well, effective 
doses of Antimony. It was found that if this drug was administered 
in sufficiently large doses to destroy the parasites, the animal died 
from severe kidney lesions; small doses, on the other hand, did not, 
even if administered repeatedly, have a noticeable effect on the 
parasites in the blood. 

It was noticed that on standing the ni. amino-phenyl-stibinic acid 
lost its action on trypanosomes, and caused on injection serious toxic 
symptoms. This fact was due to a decomposition of the compound 
into Aniline and Antimonic acid. 



37 * 







372 


II. p. AMINO-PHENYL-STIBINIC ACID 

After the somewhat discouraging results obtained with the use of 
///. Amino-phenyl-stibinic acid, experiments were undertaken with a 
view to ascertaining whether the p. compound was superior in its 
action to the ///. compound.* 

A. Rats. 

Rats infected with T. briuci , I. evansi, and T. gambiense were 
used in the following experiments: — 

The strain of T. gambiense , with which the experiments were 
carried out, was an especially virulent strain. It was recovered from 
a monkey at the time of its last relapse, a few days before death. 
Rats succumbed to the infection, on an average, 3-4 days after 
inoculation. 

As a routine method of treatment, after some preliminary 
experiments, the following procedure was decided upon: — 1st day, 
one injection of 0 5 c.c. of 5% solution, followed on the 3rd day by 
0*25 c.c. of 5% solution. The injections were repeated after a varying 
interval, as seen in Table III, pp. 374-375. 

This mode of treatment was found to be superior to injections of 
O' 5 c.c. of 5% solution on two subsequent days. Only a small 
percentage of the rats succumbed to the poisoning effects of Antimony , 
which took the form of a severe diarrhoea. At the post-mortem the 
mucous membrane of the intestine was markedly oedematous and 
inflamed ; the kidneys showed all the signs of an acute inflammation. 

The effect of the injection of the compound on the trypanosomes 
was very marked. The parasites disappeared usually after 12-1O 
hours. If treatment was discontinued the parasites reappeared in a 
comparatively short time, and a further injection again caused their 
prompt disappearance. Occasionally, after repeated injections the 
interval between relapses became shorter and shorter, until, finally, an 
injection of the drug had no influence at all on the parasites, and the 
animal died from trypanosomiasis. Table IV gives the details of 
these experiments. We were able to confirm Mesnil’s and Brimont’s 1 - 

* It is a very interesting observation of Khrlich’s that the w. Amino-phenyl- 
arsenic acid is markedly inferior in its therapeutic value to the />. Amino-phenvl- 
arsenic acid (Atoxyl). (Private communication.) 



373 


observation that this strain is not resistant to the drug in the same 
sense as Atoxyl resistant strains. If subinoculated into new rats, an 
injection of the Antimony compound caused a prompt disappearance 
of these parasites from the blood. 

B. Dogs. 

Experiments with dogs infected with 1 . evansi and 1 . brucei 
showed that these animals are very susceptible indeed to the toxic 
effect of the p . Antimony compound. If small doses were 
administered, the parasites did not disappear from the blood. If the 
doses were increased, the animal died in a very short time with severe 
kidney lesions. At the post-mortem, the kidneys were swollen and 
congested ; subcapsular and cortical haemorrhages were noticed. The 
urine was of a slight reddish colour, containing red blood corpuscles, 
casts, and large quantities of albumen. Table V, p. 377, gives the 
details of these experiments. 

Preliminary experiments on guinea-pigs proved that these animals 
reacted to the p. compound in the same way as to the m. compound. 

C. Monkeys. 

Monkeys infected with T. gavibietise were used for the following 
experiments. Treatment was usually begun when the infection was 
well established, and the animals presented undoubted signs of illness. 
Two monkeys were treated at a late stage of the infection, two at an 
earlier stage with the p. compound. Two others were treated with a 
combination of p. amino-phenvl-stibinic acid and Atoxyl. 

Experiment No. XV.— Cercofitliccus callitrichus, weight 2 k. 920 gm. Treat¬ 
ment was begun seventy-two days after infection. Numerous parasites were then 
found in. the blood. The animal was injected with 0*2 gm. of the f. compound. 
The parasites had disappeared by the next morning. Symptoms of Antimony 
poisoning had, however, set in : the monkey was vomiting white, slimy masses, and 
was suffering from severe diarrhoea ; the eyes being congested. The next day tremors 
were noticed all over the body ; the eyes were glassy and staring ; the mucous 
membrane of the mouth was cyanotic ; the temperature was subnormal, 95 0 ; the 
blood was dark and contained numerous leucocytes. These symptoms increased ; 
and the animal died in the evening of the following day. 

At the post-mortem, numerous subpleural haemorrhages were found ; the 
lungs were normal ; the heart pale and soft; the liver was very pale, and showed 
typical signs of a parenchymatous degeneration ; the spleen was enlarged ; the 
kidneys were pale and slightly congested ; the medullary and corfical substances 
not well defined ; the mucous membrane of the stomach was congested, and that 
of the intestines oedeinatous. 









375 






Table IV. 


Rat Experiment 362.— 

Inoculated with T. brucei , January 7. 


January 8. 

IO-15 to field 

... o-5 

c.c. 

of 5 % 

p. compou 

9* 

Numerous. 





10. 

N eg. 





11. 

Neg. 

... 0-25 

c.c. 



26. 

Neg. 





2 7- 

I to 5 fields. 

... o*5 

c.c. 



28. 

1-5 to field. 





29. 

Neg. 





February 3. 

Neg. 





4- 

1 to J film. 





5* 

6. 

1 to 20 fields. 

5 to field. 

... o*5 

c.c. 



7- 

Neg. 





8. 

Neg. 





9- 

1 to J film. 





10. 

1 to 2 fields. 





11. 

10 to field . 

... o-5 

c.c. 



12. 

1 to 20 fields. 





13- 

Neg. 





! 7- 

Neg. 





18. 

1 to i film. 





! 9* 

1 to 10 fields 

... o-5 

c.c. 

>» 


20. 

5 to field. 





22. 

15 to field . 

... o-5 

c.c. 

>> 

» 

2 3- 

15 to field . 

... o-5 

c.c. 

>» 

•» >> 

24. 

1 to i film. 





2 5* 

1 to field. 





26. 

5 to field . 

... o-5 

c.c. 

>> 


2 7- 






28. j 






March 1. j 

Increasing 





2 -l 

in number. 






3 - 



377 






37 8 


Experiment XXVI. — Macacus rhesus , weight 2k. 245 gm. Treatment was 
begun on the twentieth day after inoculation. The animal* was then in a very 
advanced stage of the disease. The face was puffy, the genitals swollen and 
oedematous. The blood count gave 1,570,000 red cells, 3,700 white cells and 
haemoglobin 55 per cent. o-i gm. of p. compound was then injected. The 
parasites disappeared about eleven hours after the injection, but the animal was 
found dead next morning in its cage. The post-mortem revealed the typical lesions 
of an advanced trypanosomiasis in monkeys. 

Two monkeys were inoculated at an earlier stage of the disease. 

Experiment XX. —Macacus rhesus , weight 2 k. 540 gm., was injected on the 
fifteenth day after inoculation with o-i gm. of p. compound ; a relapse set in nine 
days after the injection, when the same dose was repeated. Thirty-two days 
afterwards, a third injection of o-i gm. of the drug was administered, and then 
treatment was discontinued. The animal is still alive on the ninety-eighth day 
after inoculation, and has increased in weight to 2 k. 720 gm. The period of 
observation is, however, far too short to consider this animal cured. 

Experiment XXIV.— Macacus rhesus , weight 1 k. 650 gm. Injected with 
T. gambicnse. Very soon the animal showed oedema of the eyelids and oedematous 
swelling of the genitals. Treatment was begun thirteen days afterwards with an 
injection of o-i gm. of the p. compound. The parasites had disappeared from the 
peripheral circulation by the next day. The same dose was repeated on the 
twenty-first and thirty-seventh day after inoculation'; the treatment was then 
discontinued. The animal is still alive, sixty-four days after inoculation, and has 
increased in weight to 1 k. 720 gm. 

In order to ascertain the value of a combined Antimony-Atoxyl 
treatment in monkeys infected with T. gambiense , two monkeys 
(Macacus rhesus) were used in the following experiments : — 

Experiment XXII.— Macacus rhesus , weight 2 k. 190 gm. The animal was 
injected twenty Three days after inoculation with o*i gm. of p. compound. The 
parasites disappeared promptly from the peripheral circulation. This was followed 
on the thirty-first day by an injection of o-i gm. of Atoxyl. On the forty-eighth day 
the injection of o*i gm. of the p. compound was repeated. The animal is still 
alive, sixty-two days after inoculation, and has regained its original weight. 

Experiment XXV. —Macacus rhesus , weight 1 k. 985 gm. It was treated in the 
same way as in Experiment XXII. This animal is still alive, but in both cases 
the observation time is far too short to pronounce the animals cured. 

CONCLUSIONS 

1. The foregoing experiments prove that p. and m . amino-phenyl- 
stibinic acids are fairly powerful trypanocides, although their action is 
not so rapid as that of Sodium-antimonyl-tartrate. 

2. That the p. amino-phenyl-stibinic acid is decidedly superior 
in its action to the m. amino-phenyl-stibinic acid. 

3. Considering the satisfactory results obtained in experimental 
animals, a trial of the p . amino-phenyl-stibinic acid in patients 
suffering from Sleeping Sickness is justifiable. 

4. In our opinioa p. amino-phenyl-stibinic acid may be 
administered in the same doses as Atoxyl. As kidney lesions are 
among the most pronounced results of Antimony poisoning a careful 
systematic examination of the urine is advisable. 



379 


APPENDIX 


For the preparation of p. and m. amino-phenyl-stibinic acid we 
used the method previously adopted by Michaelis 13 for the 
production of Di-methyl-amino-phenyl-arsenic acid, employing, 
however, antimony trichloride instead of arsenic trichloride. 

Antimony trichloride was treated with aniline, and the compound 
H a N. C 6 H 5 . SbCl 2 obtained, which changed in the presence of alkali 
into the corresponding hydroxide. This formed, on treatment with 
hydrogen peroxide in an alkaline solution, aryl-stibinic acid. The 
process is expressed chemically as follows: — 

(I) H a N • C a H a + SbCl a = H a N • C 6 H 4 . SbCl* + HC 1 

(II) H*N • C„H 4 • SbCl, + 2 KOH = H„N • C e H 4 • Sb^JJ 

= H a N • C„H 4 • SbO • + H ,0 

(III) HjN • C„H 4 • SbO + H„ 0 4 = H S N ■ C,H 4 • SbO<°£ 


It is a remarkable fact that on adding aniline to melting antimony 
trichloride, p. amino-phenyl-stibinic acid (I) is formed, while, on the 
other hand, if antimony trichloride is added to boiling aniline, 
m. amino-phenyl-stibinic acid (II) is obtained. 


I 


OH 


S . b0 <OH 



NH, 



0 SbO< SU 

NH S 


For the preparation of o. amino-phenyl-stibinic acid (HI) we used 
Grignars reaction, treating o. chloraniline with antimonic acid. 


Preparation of p. amino-phenyl-stibinic acid. 

Thirty grammes of antimony trichloride are heated to 205° in a 
carefully dried flask for about 6-10 minutes, and 30 grammes of 
aniline added in three portions. The mixture is kept boiling for 
15 minutes and then poured into 500 c.c. of water. The precipitate 
which is formed is collected on a filter paper and then added to a hot 
solution of sodium carbonate, which dissolves free antimonic acid. It 
is then boiled with 80 c.c. of strong potassium hydrate (25%) for three 



hours, diluted, and 75 c.c. of commercial hydrogen peroxide added. At 
the end of two days the precipitate is boiled for two hours^ and 
filtered. On acidifying with diluted sulphuric acid; the p. amino- 
phenyl-stibinic acid is then obtained, which crystallizes in small 
needles from alcohol and water (1:3) and readily forms a sodium salt. 
The latter does not melt under 360° C., but turns brown at 323 0 C.- 
326° C. When treated with potassium iodide and sulphuric acid it is 
easily converted into p. iodo-aniline (M.P. 61 0 C.). 

Below is an analysis of the sodium salt. 

(C f H 4 0 8 NSbNa) 



Calculated 

Found 

(2) 

c 

25*26 per cent. 

24*37 P er cent. 

25-17 per cent. 

H 

2*80 

3*04 

2-92 

N 

4*91 

5 -i 8 

5 0+ 

Sb 

4 2 ’ 11 

4263 

4243 

Na 

806 

8*19 

8-27 


Preparation of m. amino-phenyl-stibinic acid. 

Twelve grammes of aniline are kept boiling, using an air 
condenser, and fifteen grammes of antimony trichloride are added in 
small portions. After all the antimony trichloride is dissolved, the 
temperature is kept up to 165° for three hours, and then the mixture 
poured into 500 c.cm. of water. The further procedure is the same 
as for the preparation of p. amino-phenyl-stibinic acid. The acid 
crystallizes from alcohol in long needles and melts at 207° C.-208 0 C 
When treated with potassium iodide and sulphuric acid it forms, 
though very sparingly, m. iodo-aniline (M.P. 26° C.). 

It is easily acetylated on boiling with acetic anhydride for two 
hours, and yields small needles which crystallize from diluted alcohol 
(1 : 15). M.P. 186° C.-188 0 C. 

Below is an analysis of the sodium salt of ;//. amino-phenyl-stibinic 
acid: 

(C.H 4 0 3 NSbNa) 



Calculated 

Found 

/ ^ 

c 

25*26 per cent. 

1 * ' 

25 03 per cent. 

(2 1 

H 

2 80 „ 

3 *i 2 

— 

N 


472 

— 

Sb 

42*11 

4276 

42 26 per cent. 

Na 

8-o6 

8-14 

819 



3 8 * 

Preparation of o . amino-phenyl-stibinic acid. 

Two grammes of o. chlor-aniline are dissolved in 30 c.c. of dry 
ether, and one gramme of antimonic acid added. After carefully 
drying for two hours at 200° C., 0*4 gramme of magnesium is added. 
The temperature rises to 45 0 C.. and is kept at this temperature for 
i-i£ hours. After evaporation of the ether, the residue is shaken up 
with 25 c.c. of sodium carbonate solution (5%), and the mixture is then 
warmed on a steam bath for one hour and filtered. The filtrate is 
acidified with diluted sulphuric acid, and the resulting precipitate 
dried and extracted with absolute alcohol. On evaporation of the 
alcohol, 0. amino-phenyl-stibinic acid is left, which crystallizes from a 
mixture of alcohol and pyridine (1 13) in leaflets. The 0. amino- 
phenyl-stibinic acid melts at I92°C. to I94°C., and forms an acetyl 
derivative on boiling with acetic anhydride (M.P. 167° C.-i6g°C.). On 
treatment with potassium iodide and sulphuric acid it yields 0. iodo- 
aniline (M.P. 54 0 C.). 


(C 6 H 4 O t N8bNa) 



Calculated 

Found 

c 

25*26 per cent. 

2512 percent. 

H 

2*80 

2*93 

N 

4-91 

516 

Sb 

4 2 ' 11 

4276 

Na 

8-o6 

823 „ 


cc 



LITERATURE 


1. Plimmer and Thomson. Further results of the experimental treatment of 

Trypanosomiasis in rats. Proc. of the Roy. Soc., Series B, Yol. So, 190S. 

2. Plimmer and Bateman. Further results of the experimental treatment of 

Trypanosomiasis. Proc. of the Roy. Soc., Series B, Vol. 80, 190S. 

3. Mesnil et Brimont. Sur Paction de Pemetique dans les Trypanosomiases. 

Bull, de la Soc. de path. exot. Tome I, No. 1, page 44. 

4. Boyce and Breinl. Atoxyl and Trypanosomiasis. Annals of Trop. Med. and 

Parasit. Vol. II, No. 1, page 32. 

5. Manson. My experience of Trypanosomiasis in Europeans and its treatment 

by Atoxyl and other drugs. Annals of Trop. Med. and Parasit. Vol. II, 
No. 1, page 46. 

6. Mesnil et Brimont. Sur la valeur curative de Pemetique dans les diverses 

Trypanosomiases. Bull, de la Soc. de Path, exot., Tome I, page 212. 

7 (<z). Laveran. Discussion, ibid., p. 216. 

(£). Laveran. De l’emploi de Pemetique dans le traitement des Trypano¬ 
somiases. Compt. rend, de l’Acad. des Scien. Tome 147, p. 510, 1908. 

8. Uhlenhuth und Woithe. Experimentelle Untersuchungen iiber Dourine 

mit besonderer Beriicksichtignug der Atoxylbehandlung. Arb. a. d. Kaisl. 
Gesundheitsamte Bd. XXIX, Heft 2. 

9. Broden and Rodhain. Travaux du laboratoire medical de Leopoldville, III, 

1907 k 1908. 

10. Martin et Darre. Trypanosomiase chez le blanc. Bull, de la Soc. de Path. 

exot.. Tome I, p. 569, 1908. 

11. Salmon. L’antimonie dans les spirilloses pathog&nes. Bull, de la Soc. de 

Path, exot., Tomme I, p. 613, 1908. 

12. Mesnil et Brimont. Sur les propri^tes des races de Trypanosomes 

r^sistantes aux medicaments. Annales Pasteur, p. 856, 1908. 

13. Michaelis. Uber die p. di-methyl-amino-phenyl-arsinsaure (di-methyl Atoxyl i. 

Ber. d. deutsch. chem. Gesell., Vol. 41, p. 1514 (1908). 



SHORT NOTE ON THE MECHANISM OF 
HAEMOLYSIS IN PIROPLASMOSIS 

CANIS 


BY 

ANTON BREINL, M.U.Dr., 

DIRECTOR, RUNCORN RESEARCH LABORATORIES, LIVERPOOL SCHOOL OK 
TROPICAL MEDICINE 

AND 

H. E. ANNETT, M.D., D.P.H., 

PROFESSOR OF COMPARATIVE PATHOLOGY, UNIVERSITY OF LIVERPOOL 

(Received for publication 0 April , 1909^ 

For the following researches puppies and dogs of various ages 
were infected with our laboratory strain of Piroplasma canis. The 
infection was invariably fatal for young puppies up to the age of 
2 \ months. After a varying incubation period, the animals shewed 
parasites in the blood in small numbers at first, but very soon a rapid 
multiplication occurred and lasted for 24 to 36 hours, accompanied by 
haemoglobinuria. The urine was of a light reddish to dark mahogany 
colour, and the more rapid the multiplication of the parasites, the 
darker was the colour of the urine. Pronounced jaundice of the 
tissues was only noticed in two out of seventy animals used. In these 
young animals the blood serum, taken at death, was of a dark reddish 
to mahogany colour, according to the colour of urine. 

In dogs and older puppies, the disease was hardly ever fatal. The 
parasites did not multiply so rapidly, and nearly always were present 
in smaller numbers; we have never observed a pronounced 
haemoglobinuria in these animals, but for three weeks after inoculation 
the serum was slightly reddish in colour. 

It was especially noticed that in young puppies a rapid diminution 
of the blood corpuscles took place during the last few hours before 
death. 

It seemed, therefore, of interest to determine whether the 
destruction of the blood corpuscles was due to Isolysins or Autolysins. 
With this object, the serum of infected animals in different stages of 
the disease was added in varying dilutions to red blood corpuscles of 



.*84 

the infected animals from which the serum was obtained, and also to 
blood corpuscles of normal dogs or puppies. 

The serum was used both fresh and after being heated for 
thirty minutes at a temperature of 56° C.; and in this latter case with 
and without the addition of complement (fresh centrifugalised dog 
serum). 

The blood corpuscles were usually washed three times in 0*9% 
saline solution, and used in a 10% suspension. 

Serum of a dark reddish colour taken from a puppy three hours 
before death : haemoglobinuria marked. 

c.( . r.c. r.c. c.c. c.c. c.c. 

A. 10 per cent, suspension of 
infected dog’s red blood 

corpuscles ... ... 1 1 1 1 1 1 


Fresh normal dog’s 
(diluted 1 in 10) 

serum 

1 

1 

1 

I 

I 

I 

Infected dog’s inactivated 







serum 

... 

1 

° - 5 

O'1 

005 

001 

0005 

Two hours at 37 0 C. 

... 

Slight 

Haemo¬ 

lysis 

0 

0 

0 

0 

0 

10 per cent, suspension of 
infected dog’s blood 
corpuscles . 

I 

I 


I 

I 

I 

Infected dog’s serum 


I 

05 

O'I 

005 

O'OI 

0005 

Two hours at 37 0 C. 


Slight 

Haemo¬ 

lysis 

0 

0 

0 

0 

0 

10 per cent, suspension of 
infected dog’s blood 
corpuscles 


1 

I 

I 

I 


Normal serum 


I 


O'I 

005 

O'OI 

0005 

Two hours at 37 0 C. 


O 

0 

0 

0 

0 

0 


D. 10 per cent, suspension 
of normal dog’s blood 
corpuscles ... ... 1 

Fresh normal dog’s serum 
(diluted 1 in 10) ... 1 

Inactivated infected dog’s 
serum ... ... ... 1 

Two hours at 37 0 C. ... o 


111 11 

111 1 1 

o*5 o* 1 005 o*oi 0005 

0000 o 



J«5 


c.c. c.c. c.c. c.c. c.c. c.c. 

E. io per cent, suspension 
of normal dog’s blood 

corpuscles . i i i i i i 

Infected dog’s serum ... i 0*5 01 0*05 o*oi 0 005 

Two hours at 37 0 C. ...0000 o o 

Similar experiments were carried out with the serum of a larger 
dog, withdrawn during the early acute stage of the infection, when 
parasites were present in small numbers. The serum was of a light 
reddish colour. The results were entirely negative : in none of the 
test tubes was there any sign of haemolysis. 

The series of experiments was repeated in the same way as in the 
first experiment, with the serum of a puppy; the serum of which was 
of a dark brownish mahogany colour; and also in these experiments, 
1 c.c. of the infected serum added to 1 c.c. of 10% infected blood 
corpuscles, with both activated and inactivated serum, caused an 
extremely slight haemolysis, whereas in no other tube was there any 
sign of haemolysis noticed. 

The same experiment was repeated with the serum of a dog three 
weeks after the first appearance of the parasites, and also with the 
serum of a puppy at the commencement of the rise of temperature. 
In no case was there any haemolysis of either normal red blood 
corpuscles, or of the red corpuscles of the infected animal from which 
the serum was obtained. 


CONCLUSION 

These experiments tend to prove that the haemolysis and the 
haemoglobinuria in infections with Piroplasma canis is due neither 
to an Isolysin nor to an Autolysin, but apparently only to a 
disintegration of red blood corpuscles after the escape of the parasites 
from them. 




387 


GLAND PUNCTURE IN THE DIAGNOSIS 
OF ANIMAL TRYPANOSOMIASIS 

BY 

R. EUSTACE MONTGOMERY, M.R.C.V.S., 

AND 

ALLAN KINGHORN, M.B., 


(Received for publication 15 Apr if 1909,) 


Acting on a suggestion made by Mott, Greig and Gray examined 
the juice obtained by puncture of the enlarged lymphatic glands of 
men suffering from trypanosomiasis in Uganda. Dutton and Todd 
in the Congo Free State also noted their connection with the disease, 
and instituted a system of census dependent upon their size. They 
found that 91% of natives having post-cervical glands measuring 
approximately 1*5 by 075 cm. showed trypanosomes on puncture. 

Enlargement of the superficial lymphatic glands in animals, 
though well recognised post-mortem, has not received much 
attention as a symptom of trypanosomiasis. Dutton, Todd and 
Kinghorn* quote Bertolloti as having noted it to be a constant feature 
in the infected stock at Eala; and enlargement of the presternals is 
well known to camel owners in the Punjab, and by some at least is 
associated with Surra. In Rhodesia we found it to be common in all 
classes of animals sick and healthy, even young calves and several 
varieties of antelope having almost without exception easily palpable 
glands. As a symptom, then, it is here of little value. 

During the time one of us (R. E. M.) was in India, a few 
observations were made as to the value of gland puncture in camels 
believed to be suffering from Surra, but not showing trypanosomes in 
the peripheral circulation. In one case a camel which did not show 
peripheral organisms for sixty-three consecutive days revealed them 
on three out of four punctures ; and on another occasion when 
examining a herd, two additional cases were discovered by this 
method. 


* Annals of Tropical Medicine and Parasitology, 11)07, Yol. 1, p 235. 




Our observations on cattle suffering from T . dimorphon at 
Broken Hill* did not accord with those of Dutton and Todd on 
Congolese cattle. The infection in our animals, however, was acute, 
and trypanosomes were rarely absent from the circulation: in such 
cases it is unnecessary to resort to gland puncture. Our more recent 
observations on animals suffering from a somewhat chronic form of 
disease leads us to regard it as a most valuable diagnostic method. 
It is not only of value in chronic cases, but should animal inoculation 
have been carried out, a positive examination may be obtained some 
days prior to the appearance of trypanosomes in the blood. We may 
quote the following cases: — 

A goat inoculated with a cattle strain of T. dimorphon showed trypanosomes 
on gland puncture from the ninth day: they only appeared in the blood (i in 
J 4 cover-glass) on the fourteenth. 

A dog inoculated with the same strain never showed peripheral trypanosomes 
up to death on the thirty-fifth day. Gland puncture on the tenth and nineteenth 
days was positive. 

An ox inoculated with a trypanosome allied to T. brutei showed organisms in 
the prescapular glands two days prior to their appearance in the blood. 

An ox inoculated with a trypanosome of doubtful nature (Ninamwenda strain) 
showed organisms in the gland three days before they were seen in the blood. 

A dog inoculated with a dog strain of tadpole trypanosomes showed organisms 
on gland puncture eleven days previous to their being seen in the blood. 

Infections of the goat due to T. brucei and T. pecaudi are known 
to be easily overlooked, unless means of sub-inoculation are at hand. 
The following cases are instances of the advantages of puncture : — 

A goat inoculated with a form allied to T. brucei, which was under observation 
for six weeks, only showed trypanosomes on two occasions. They were present on 
each of five gland punctures when not seen in the blood. 

A goat inoculated with the same strain did not show blood trypanosomes during 
the fortnight available for observation. Gland puncture showed them present 
from the seventh day. 

It is, of course, hardly to be expected that success is invariable. 
In animals known to be infected we have often been unable to detect 
their presence on puncture; but the advantages of even one positive 
case fully compensate for the small amount of labour involved in 
adopting this method as a routine for all suspected animals not 
showing peripheral trypanosomes. 

The advantages in practice may be judged by the fact that in 
three localities fifteen cases were diagnosed on blood examination, 
and four additional ones on puncture; whilst at another place. 


* Armais of Tropical Medicine and Parasitology, 1908, Vol. 2, p. 106. 



considered to be free of disease, it was by gland puncture alone that 
its existence was demonstrated. 

We are indebted to Mr. Lane, the Veterinary Officer of North 
Eastern Rhodesia, for the following summary of results since adopting 
this method of diagnosis. The trypanosome with which he had 
chiefly to do is, in animal reaction and morphology, related to 
T. natium. On one Station four cases were diagnosed by gland 
puncture : in no instance were trypanosomes seen in the blood. On a 
second Station, in five cases the gland juice was positive, while only 
two showed organisms in the blood. 




>" ■ 

OBSERVATIONS ON THE HOOKLETS OF 
CTSTICERCUS CEUJJLOSAE IN MAN 

j w vv. blkfUKNS. \{ ( A.m I) V I) . 

V \> • •• •/ >? C. M m, {i «« (v* 

•(■fifr.ai-tH i : Q? pupil* ah>ji 3 ; i Apf U\ O'KH v 

fa Miyy.igiA, Dr> < '..iiapbeH and ftn.^uu; *A taiuiiit'Otfiriiidagu., 
Mv)|lr;r- kl-njK MH'-'-hif'C) p }Uv limsrum -,iM ; !,mvu ••! i i'S {t‘ r f ' '7 • 
v//,:-: in m [.ivr liYu-di' ••• Vi,, i *. \ irir ~./v • >i' ?b> : 







tissue capsules of the cy' 3 ts ; varied from 15-21 mm. long by 8-10 mm. 
broad (fig. i). Recently 1 proceeded to examine a scolex extracted 
ft opt its bjiidder noth a view to making certain of the diagnosis. I was 
-luprised .icrorduigly, on exanviimg a specimen, to find only sixteen 
booklets instead of twenty-two to thirty-two, which is the . number 
given by various, authorities .as comprising the limits of variation. It 
was possible that one circle, or booklets was absent, but on measuring, 
this explanation, taken also m connection with what will appear later, 
is hardly possible. 


Kit-, Ctrdf *‘»i booklets t>i the same, x tit* jappr<toij#*teJy 

1. i'GvljM!' cysih-trt.cs (hg. 2). Sixteen booklets,* The size of the 
hooks varied from to 8 x*,?> i4«l’0/t As wifi be seen from the 
appended protocols, there was no sharp demarcation between small 
and large hooks, but hootdHs of various str.es also occurred,- £#., 

1 $>'*}/*. ! 20 t>/‘. * .1 *•' *•“ V ! ■ 

>. . Put oral cy.<(n-<r.<tfS-. Twenty-one booklets found. The range 
of variation was in this case greater, viz., from 104*4-t-*2'4.u for vyhat 
jvi.ght be galled sniali iiuoiv-., and from (qp'S-if-yCt« for the large. 







3- Pectoral cy s lie er cits. Booklets twenty Only si few -hooks 
were measured, three small, varying from roS'CM rrb.u, and five Satg^feV 
varying from i44-0-; 51 •*.**. 

4. Pea oral cysfpenus. TVentv booklets. In this case, as m 
case No, 1 , it is hardly possible to separate the hooks into a large and 
srpai) senes, ns hooks of an intermediate sue occur, Thus hooks of 
the following, -sixes--' were measured 122-5, cso c. (jvo, 140, 14-3*5, 
■ 47 '°* iW v 153-5/0 

f next examined a specimen of i'PcUUiiasac from the brain add 4 
specimen ftbni the tongue , both frohi hatiyes of Madras and presented 


lit. > ~ ( I iirenu ? rplluh <' l ? fYom I'Hiill X 80 Uj>|d'O.ximatdV 


to the museum i;«y jSi'ajur' 'W-ilhaais, 1 ,Ai,S>, s and..compared them wilt, 
the hooks of || udmm 10 man a* a) c. t .//;,<An «>■ from the pig. 

5. fn-.-mt cvw/t.cfiH.y (fig, 3). id00k lets nv<-m\ .eight. The small ' 
range from so> o fs8'8p ; the Marge "trod. 151 ?• 1O2 o/u. s<> that there 
m .a fairly well .market! line of sepc-rawa, 

O. Ti'ttgve mm'c-e/. ;/>'(% -p- One month's? dufatioti. llookk;;- 
twenty-two f ? two missing). The small range Iron. foibo-i^w 
The large from t ,}• > 4 15t '2». The range of variation is riot so great. 





m H I 


m.i >•, Hi- Imr •<( ration‘the amdi and large sr< marked 

<is< in the brain cvs-tirefcus. 

-Jb Pig- -tfsksch vyiiU(rtiis. '■ . jfo'etilyd&v* booklet* found. Ten 
hdokleis yvefr- measured- The -sire 6 f the small was constant, viz., 
1 i’t’/i. That of the large >.vas also 1 /instant, u?,;, 17^ft, so that 
separation betvveen large arid strttbi w,is. qhite. distmct:, 




fto appr^timattlv 


to. i so/mw. Twentyffive: kf«rk.teN : 'v*mte! ;feitt)di‘' Eighteen of 
these were measured. The sm&ii yangt:from Hya-i^rr^p and the 
large from iH.Vt- itoy?/*, so that, the line of separation - is again 
d 1-'ththough it is noticeable that the size of the booklets is 
distinctly-larger uian m the case oi. the cysncercus in the pigs muscle. 

it y<.httlcl ■‘jkpif^’dr, therefore; from these-.observation's that in 
!. . (■t/fiihun it; Utah there is ;ui irregularity of development affecting- 
both the number of the booklets and. more especially, their size. 




I C. cellulosae fron 

solium Brain cyst. Tongue cyst j Pectoral cv^r i. Pectoral cyst 2 Pectoral cyst 3. Pectoral cyst 4. pig's muscle. 


*-> t\(M m r^.(N 


■ r^o© 3" — n m -t-vo r%x x | -- v 

n - r 


O O VC O O O M fl 


+ o c x y. m m */. oe oc m x m co 00 +0 O o 77 

-f zb - M X 73 ri ^ "t-vo 

O O x » - » f| in in in i/% i/-. mi 1/-1 tD »o • r. C vC VC V O 


— ci f*", i- >no rv oc cv o - ri co -1- «-ovc r^oo 


G VC VC VC VC VC vO N Cl r| r| GO OC 0 

c/3 vc vc vc vc jv ov n f’n r<i MC 6 *j- 

O N M fl Cl rj rj r, n M CO ro «+■ 


►x d CO -h tn\C OO cv O — N co-t-i/l 


N O O O '£ C fl N N ri */) Cp + -+• O p vC vO O >C vC vO cl | 7 7 

« X OO - '*ci in in iV.co OO fl O -f t K N N N l\ fv ■- i O -f 

-1- -t- 1- 1- -h ^ ^ ^ o 


— N «-o -f «ov,O fvoO O O 


vCOOOOOOOvCNMfioooo flXXXSOM + + 0 O 


MSC 5 C 7 I- 71 - 7 I- w ‘-P Cf 7 + 
•'o®5 CC Cl cl c| iV. sC VC b 


. rv .. • • I ••••••••] 

P- M - f| fC-t'/-.c K/J J^O I — Cl CO - 1 - 10VC t^oo _J 














































Volume II 


May, 1909 


No. 5 


ANNALS 

OF 

TROPICAL MEDICINE AND 
PARASITOLOGY 


ISSUED BY 

THE LIVERPOOL SCHOOL OF TROPICAL MEDICINE 


Editor 

Professor RONALD ROSS, Major I.M.S. (Ret.), D.P.H., F.R.C.S., 
D.Sc., LL.D., F.R.S., C.B. 


In Collaboration with 

J. W. W. STEPHENS, M.D., Cantab., D.P.H. 

R. NEWSTEAD, M.Sc., A.L.S., F.E.S., Hon. F.R.H.S. 
J. L. TODD, B.A., M.D., C.M. McGill, D.Sc., Liv. 
H. WOLFERSTAN THOMAS, M.D., C.M. McGill. 
ANTON BREINL, M.U.Dr. 

Professor Sir RUBERT BOYCE, M.B., F.R.S. 



( \ Tinting fir- Co., Tt.i. 

Printers to the University Press of Z.i-r,er/>, 
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r><» 



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COMMITTEE 

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Professor - - MAJOR RONALD ROSS, F.R.C.S., D.P.H., D.Sc., 

LL.D., F.R.S., C.B. Nobel Laureate 1902. 
(Indian Medical Service, retired). Sir Alfred 
fones Professor of 7 'ropical Medicine 

Lecturers - JOHN WILLIAM WATSON STEPHENS, M.D.. 

Cantab., D.P.H. Walter Myers Lecturer 
ROBERT NEWSTEAD. M.Sc., A.L.S., F.E.S. 
Lecturer on Medical Entomology> 

Assistant Lecturers - HAROLD WOLFERSTAN THOMAS, M.D., 
C.M. 

ANTON BREINL, M.U.Dr. 

R. STENHOUSE WILLIAMS. M.B., D.P.H., 
Assistant Lecturer in Public Health Bacteriology 

Honorary Lecturers - WILLIAM THOMAS PROUT, M.B , C.M.G. 

Major JOSEPH FITZGERALD BLOOD, M.D., 
M.Ch. (Indian Medical Service, retired) 

2. At the Royal Southern Hospital, Liverpool 

Physicians - - CHARLES JOHN MAC A LISTER, M.D., 

M.R.C.P. 

JOHN LLOYD ROBERTS, M.D.. M.R.C.P. 
Major RONALD ROSS, C.B.. F.R.S.. etc. 

Surgeons - - WILLIAM ALEXANDER, M.D., F.R.C.S. 

ROBERT JONES, F.R.C.S. 

GEORGE PALMERSTON NKWBOLT. M B., 
F.R.C.S. 

Tropical Pathologist - JOHN WILLIAM WATSON STEPHENS, M.I). 

3. At the Runcorn Research Laboratories 

Director - - - ANTON BREINL, M.U.Dr., Pia^. 

R TdLh orators } M ‘ NIERENSTEIN. Ph.D., and other workers 

4. On Expeditions 

HAROLD WOLFERSTAN THOMAS, M.D., 
CM. 

ALLAN KINGHORN, M.B. 

ROBERT EUSTACE MONTGOMERY, 
M.R.C.V.S. 

JOHN OGLETHORPE WAKELIN BARRATT, 
M.D., D.Sc. (Lond.) 

WARRINGTON YORKE, M.D. 

Dean 

Professor Sir RUBERT BOYCE, M.B., F.R.S. 

Laboratory 

JOHNSTON TROPICAL LABORATORY, LIVERPOOL 
UNIVERSITY 

Research Laboratory 
CROFTON LODGE, RUNCORN 
Hospital 

ROYAL SOUTHERN HOSPITAL. LIVERPOOL 
Secretary*>'s Office 

B 10, EXCHANGE BUILDINGS, LIVERPOOL 



NOTICE 


The following changes in the courses of instruction given by the 
Liverpool School of Tropical Medicine will come into force in and 
after January, 1909: — 

Full Course begins 6 January. Short Course begins 1 June. 
Diploma Examination, 5 April. Certificate Examination, 29 June. 
Full Course begins 15 September. 

Diploma Examination, 13 December. 

The full Course of Instruction is open to all qualified medical men, 
and the examination to all students who have taken out this full 
course. 

Fee for the full Course of Instruction—Thirteen guineas. 

Fee for the Diploma Examination—Five guineas. 

Fee for the Short Course of Instruction—Four guineas. 

Fee for the use of a School microscope during one term—Ten 
shillings and sixpence. 

For prospectus and further information, application should be made 
to the Dean of the Medical Faculty, University of Liverpool 


The following have obtained the Diploma in Tropical Medicine of 
the University of Liverpool: — 

Diploma in Tropical Medicine 


Date of 
Difloma 

1909 Abercrombie, Rudolph George 

1906 Adie, Joseph Rosamond 

1907 Allan, Alexander Smith 
1909 Allin, John Richard Percy 
1907 All wood, James Aldred 

1905 Anderson, Catherine Elmslie 

1906 Arnold, Frank Arthur 
1904 Augustine, Henry Joshua 
1909 Barrow, Harold Percy Waller 
1906 Bate, John Brabant 

1904 Bennett, Arthur King 

1906 Bennetts, Harold Graves 

1907 Bond, Ashton 

1907 Branch, Stanley 

1905 Brown, Alexander 
1904 Bruce, William James 

1904 Byrne, John Scott 

1905 Caldwell, Thomas Cathcart 
1909 Carr-White, Percy 

1906 Carter, Robert Markham 

1908 Caverhill, Austin Mack 
1906 Chisholm, James Alexander 

1909 Clark, William Scott 

1904 Clayton, Thomas Morrison 

1906 Clements, Robert William 

1907 Collinson, Walter Julius 


Date of 
Difloma 

1909 Cope, Ricardo 

1908 Crawford, Gilbert Stewart 

1905 Critien, Attilio 

1908 Dalai, Kaikhusroo Rustomji 
1904 Dalziel, John McEwen 
1908 Dansey-Browning, George 

1907 Davey, John Bernard 

1908 Davidson, James 
1904 Dee, Peter 

1908 Dickson, John Rhodes 

1907 Donaldson, Anson Scott 

1908 Dowdall, Arthur Melville 

1906 Dundas, James 

1906 Faichnie, Norman 

1907 FeU, Matthew Henry Gregson 

1907 Gann, Thomas William Francis 

1908 Glover, Henry Joseph 

1907 Graham, James Drummond 

1908 Greaves, Francis Wood 
1904 Greenidge, Oliver Campbell 
1908 Goodbody, Cecil Maurice 

1908 Harrison, James Herbert Hugh 

1909 Hayward, William Davey 

1904 Hehir, Patrick 

1907 Hiscock, Robert Carroll 

1905 Hooton, Alfred 



Date of 
Diploma 

1905 Hudson, Charles Tilson 

1905 Illington, Edmund Moritz 

1906 Jeffreys, Herbert Castelman 
1908 Joshi, Lemuel Lucas 

1907 Keane, Joseph Gerald 
1907 Kennan, Richard Henry 

1907 Kenrick, William Hamilton 
1904 Khan, Saiduzzafor 

1904 Laurie, Robert 

1908 Le Fanu, Cecil Vivian 

1907 Le Fanu, George Ernest Hugh 

1908 Luethgen, Carl Wilhelm Ludwig 

1905 Macfarlane, Robert Maxwell 

1906 Mackenzie, Donald Francis 

1907 Mackey, Charles 

1904 Maclurkin, Alfred Robert 

1905 Maddock, Edward Cecil Gordon 

1907 Maddox, Ralph Henry 

1908 Mama, Jamshed Ryramji 

1907 McCarthy, John McDonald 

1908 McCay, Frederick William 

1904 McConnell, Robert Ernest 

1908 McLellan, Samuel Wilson 
*909 Meldrum, William Percy 

1905 Moore, James Jackson 

1909 Murphy, John Cullinan 

1904 Nicholson, James Edward 

1905 Nightingale, Samuel Shore 

1906 Pailthorpe, Mary Elizabeth 


Date of 
Diploma 

1906 Palmer, Harold Thornbury 

1908 Pearce, Charles Ross 

1906 Pearse, Albert 

1904 Philipson, Nicholas 

1905 Radcliffe, Percy Alexander Hurst 

1907 Raikes, Cuthbert Taunton 

1907 Ryan, Joseph Charles 

1906 Sampey, Alexander William 

1909 Samuel, Mysore Gnananandaraju 

1908 Scboorel, Alexander Frederik 
1904 Sharman, Eric Harding 
1908 Smith, John Macgregor 
1906 Smithson, Arthur Ernest 
1908 Stewart, George E tward 

1908 Tate, Gerald William 

1906 Taylor, Joseph van Someron 
1906 Taylor, William Irwin 
1904 Thomson, Frank Wyville 

1909 Thornely, Michael Harris 

1906 Tynan, Edward Joseph 

1907 Vallance, Hugh 

1904 Walker, George Francis Clegg 
1906 Watson, Cecil Francis 

1909 Webb, William Spinks 

1908 Whyte, Robert 

1906 Willcocks, Roger Durant 
1906 Williamson, George Alexander 

1905 Young, John Cameron 



EDITORIAL NOTICE 


By order of the Committee of the Incorporated Liverpool School 
of Tropical Medicine, the series of the Reports of the School, which 
have been issued since 1899, will be followed, from January 1, 19° 7 » 
by the Annals of Tropical Medicine and Parasitology, of which 
this is the fifth number of the second volume. 

The Annals are issued by the Committee of the School, and will 
contain all such matter as was formerly printed in the Reports—that 
is to say, accounts of the various expeditions of the School and of the 
scientific work done in its laboratories at the University of Liverpool 
and at Runcorn. Altogether twenty-one Memoirs, besides other 
works, have been published by the School since 1899, and of these 
ten, containing 519 quarto or octavo pages and 95 plates and figures, 
were published during the two years 1904 and 1905 ; and there is no 
reason to suppose that this rate of production by the workers of the 
School alone will diminish in the future. In addition, however, to 
School work, original articles from outside on any subject connected 
with Tropical Medicine or Hygiene may be published if found 
suitable (see notice on back of cover); so that, in all probability, not 
less than four numbers of the Annals will be issued annually. Each 
number will be brought out when material sufficient for it has been 
accumulated. 




THE UNIVERSITY PRESS OF LIVERPOOL 


Publications of the 

Liverpool School of Tropical Medicine 

MEMOIR i 

Malarial Fever: its Cause, Prevention, and Treatment (1903). Con¬ 
taining full Details for the use of Travellers, Sportsmen, Soldiers, and Residents in 
Malarious Places. By Ronald Ross, C.B., F.R.S., F.R.C.S. With Frontispiece 
and Plate. 8vo. 2s. 6d. 

MEMOIR II 

Report of the Malaria Expedition to Sierra Leone (1899). By 
Ronald Ross, D.P.H., M.R.C.S., H. E. Annett, M.B., D.P.H., and E. E. Austen. 
Being a full account of the first expedition of the School, and containing besides 
much matter relating to the parasites of malaria, to the gnats which carry them, and 
embodying some previous observations of Major Ross in India. Illustrated by four 
maps and five full-page collotypes. Quarto. Price 21s. nett. 

MEMOIR III 

Report of the Malaria Expedition to Nigeria (1900). By H. E. Annett, 
M.D., D.P.H., the late J. Everett Dutton, M.B., Ch.B., and J. H. Elliott, M.D. 
Part 1. Malarial Fever , etc. Giving a full account of the expedition, with numerous 
views in the text, charts, maps, and two plates, and containing much matter of 
general importance. Quarto. Price 10s. 6d. nett. 

MEMOIR IV 

Report of the Malaria Expedition to Nigeria (1900). By the same 
authors. Part 11. Filariasis. Containing many new observations upon Filariae of 
Birds, with numerous illustrations and nineteen plates, five of which are coloured 
and give the microscopical anatomy of the head of Anopheles costalis (by the late 
Dr. Dutton). Quarto. Out of print. 

MEMOIR V, PART 1 

First Progress Report of the Campaign against Mosquitoes in 
Sierra Leone (1901). By Major R. Ross, F.R.C.S., D.P.H., F.R.S., dated 15th 
October, 1901, giving details of the commencement of the Campaign, with a letter 
from Dr. Daniels regarding the results arrived at to date. 8vo. Price is. 

MEMOIR V, PART 2 

Second Progress Report of the Campaign against Mosquitoes in 
Sierra Leone. By M. Logan Taylor, M.B., dated 15th September, 1902. 8vo. 
Price is. 

MEMOIR VII 

Report of the Yellow Fever Expedition to Para (1900). By H. E. 
Durham, M.B., F.R.C.S., and the late Walter Myers, M.B. (Dr. Walter Myers 
died of Yellow Fever whilst serving on this expedition). Quarto. Out of print. 


1 



MEMOIR VIII 

Report on the Sanitary Condition of Cape Coast Town (1902). 
With suggestions as to improvement of same, by M. Logan Taylor, M.B., Ch.B. 
8vo. Price is. 

MEMOIR IX. 

Report on Malaria at Ismailia and Suez (1903). By Ronald Ross, 
F.R.C.S., D.P.H., F.R.S., C.B. 8vo. Price is. 

MEMOIR X 

Report of the Malaria Expedition to the Gambia (1902). By the late 
J. E. Dutton, M.B , Ch.B. Quarto. Price 15s. nett. 

MEMOIR XI 

First Report of the Trypanosomiasis Expedition to Senegambia 
(1902). By the late J. Everett Dutton, M.B., Ch.B., Viet., and John L. Todd, 
B.A., M.D., C.M., McGill. Quarto. Price 15s. nett. 

MEMOIR XII 

The Anti-Malaria Measures at Ismailia (1904). Bv Rubert Boyce, 
M.B., F.R.S. Price is. 

MEMOIR XIII 

Reports of the Trypanosomiasis Expedition to the Congo (1903-1904). 
By the late J. Everett Dutton, M.B., John L. Todd, M.D., and Cuthbert 
Christy, M.D. Quarto. Price 21s. nett. 

MEMOIR XIV 

Report on the Sanitation and Anti-Malarial Measures in practice in 
Bathurst, Conakry, and Freetown (1905). By Rubert Boyce, M.B., F.R.S., 
Arthur Evans, M.R.C.S., H. Herbert Clarke, M.A., B.C., Cantab. Quarto. 
Eight plates. Price 5s. 

MEMOIR XV 

General Sanitation and Anti-Malarial Measures in Sekondi, the 
Goldfields, and Kumassi, and a comparison between the conditions of 
European residence in India. By Lieut.-Colonel Giles. Quarto. Price 7s. 6d. 
nett. 

MEMOIR XVI 

Trypanosomes, Trypanosomiasis, and Sleeping Sickness : Pathology 
and Treatment. By H. Wolferstan Thomas, M.D., McGill, and Anton Breinl, 
M.U.Dr., Prag. Quarto. 6 plates (5 coloured) and 7 charts. Price 12s. 6d, nett. 

Gland Puncture in Trypanosomiasis. By the late J. Everett Dutton, 
M.B., Viet., and John L. Todd, B.A., M.D., McGill. Two charts and one figure. 

MEMOIR XVII 

The Nature of Human Tick-Fever in the Eastern part of the Congo 
Free State. By the late J. Everett Dutton, M.B., and John L. Todd, B.A., 
M.D., McGill. Quarto. With map, four plates, and nine temperature charts. 
Price 7s. 6d. nett. 

On the External Anatomy of Ornithodoros moubata. By R. Newstead, 
A.L.S., F.E.S. With two plates. 

ii 



MEMOIR XVIII 

I. Gland Palpation in Human Trypanosomiasis; 

AND 

II. The Distribution and Spread of 4 Sleeping Sickness* in the Congo 
Free State. By the late J. Everett Dutton, M.B., and John L. 
Todd, B.A., M.D., C.M., McGill. With four maps (2 colours) and four 
plates. 

III. A New Dermanyssid Acarid. By R. Newstead, A.L.S., F.E.S., and 

John L. Todd, B.A., M.D., C.M., McGill. With one plate. 

IV. Another New Dermanyssid Acarid. By R. Newstead, A.L.S., F.E.S. 

With one plate. 

V. Anatomy of the Proboscis of Biting Flies. By J. W. W. Stephens, 
M.D., Cantab., and R. Newstead, A.L.S., F.E.S. With six plates. 

Imp. 8vo. Price 7s. 6d. nett. 

MEMOIR XIX 

Yellow Fever Prophylaxis in New Orleans in 1905. By Rubert 
Boyce, M.B., F.R.S. Imp. 8vo. Maps and six plates. Price 5s. nett. 

MEMOIR XX 

I. La prophylaxie de la Malaria dans les principaux postes de l’£tat 

Ind6pendant du Congo. By the late J. Everett Dutton, M.B., 
and John L. Todd, B.A., M.D. With four maps and four illustrations. 

II. The Animal Reactions of the Spirochaeta of African ‘Tick Fever.’ 

By Anton Breinl, M.D., and A. Kinghorn, M.B. 

III. The Specific Nature of the Spirochaeta of African ‘Tick Fever.’ 
By Anton Breinl, M.D. Imp. 8vo. Price 5s. nett. 

MEMOIR XXI 

I. The Runcorn Research Laboratories of the Liverpool School of 
Tropical Medicine. With five plates. 

II . An Experimental Study of the Spirochaete of the African Tick 

Fever (Spirochaeta duttoni). By Anton Breinl, M.U.Dr., and 
Allan Kinghorn, M.B. With one figure and ten charts. 

III. A Note on a New Spirochaete found in a Mouse. By the same 

authors. With one figure. 

IV. Comparison between the Trypanosomes Present by Day and by 

Night in the Peripheral Blood of Cases of Human Trypano¬ 
somiasis. By the late J. Everett Dutton, M.B., John L. Todd, 
B.A., M.D , C.M., and E. N. Tobey, A.B., A.M., M.D. 

V. The Lesions in the Lymphatic Glands in Human Trypanosomiasis. 
By R. Howard Mole, M.D. With two plates. 

VI. Concerning certain Parasitic Protozoa observed in Africa. By 
the late J. Everett Dutton, M.B., John L. Todd, B.A., M.D., C.M., 
and E. N. Tobey, A.B.,A.M., M.D. With two figures and one coloured 
plate. 

VII. Attempts to Cultivate Spirochaeta duttoni. By Lewis A. Williams, 
M.D., D.P.H., and R. Stenhouse Williams, M.B., D.P.H. 

VIII. Attempts to Transmit Spirochaetes by the Bites of Cimex 
lectularius. By Anton Breinl, M.U.Dr., Allan Kinghorn, M.B., 
and John L. Todd, B.A., M.D. Imp. 8vo. Price 7s. 6d. nett. 

iii 



ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. I. No. i. 


Insects and other Arthropoda collected in the Congo Free State. 

By R. Newstead, A.L.S., F.E.S., the late J. E. Dutton, M.B., and 
John L. Todd, B.A., M.D., C.M., McGill. Six plates. 

Description of Two New Species of African Ticks. By H. Neumann. 

On some Parasites in the Museum of the Liverpool School of Tropical 
Medicine. By A. Looss. Three plates. 

The Presence of Spirochaeta Duttoni in the Ova of Ornithodoros 
Moubata. By Capt. R. Markham Carter, I.M.S. One plate. 

A note on the Therapeutics of Trypanosomiasis. By Benjamin Moore, 
M.A., D.Sc., M. Nierenstein, Ph.D., and John L. Todd. B.A., M.D., 
C.M., McGill. Imp. 8vo. Price 7s. 6d. nett. 


ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. I. No. 2. 

An Automatic Oiler for the Destruction and Prevention of Mosquito 
Larvae in Cesspools and other collections of Water. By E. H. Ross, 
and H. C. Ross. One plate. 

The Anatomy of the Proboscis of Biting Flies. Part II, Stomoxys. By 
J. W. W. Stephens, M.D., and R. Newstead, A.L.S., F.E.S. Eight plates. 

Trypanosome Transmission Experiments. By the late J. Everett Dutton, 
M.B., John L. Todd, B.A., M.D., C.M., McGill, and J. W. Hanington. 
M.D., McGill, 

Cattle Trypanosomiasis in the Congo Free State. By the late J. Everett 
Dutton, M.B., John L. Todd, B.A., M.D., C.M., McGill, and Allan 
Kinghorn, M.B. Two plates and six charts. 

Concerning the Treatment of Experimental Trypanosomiasis. By 
Benjamin Moore, M.A., D.Sc., M. Nierenstein, Ph.D., and John L. Todd, 
B.A., M.D., C.M., McGill. Imp. 8vo. Price 7s. 6d. nett. 


IV 



ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. I. No. 3. 


Concerning certain Parasitic Protozoa observed in Africa. By the late 
J. Everett Dutton, M.B., Viet., John L. Todd, B.A., M.D. McGill, and 
E. N. Tobey, A.B., A.M., M.D. Harvard. Thirteen plates. 

Yaws. By C. W. Branch, M.B., C.M. (Edin.) 

A Description of some Gold Coast Entomostraca. By YV. M. Graham, 
B.A., M.B. 

Notes on Dr. Graham s Collection of Cyclopidae from the African Gold 
Coast. By G. Stewardson Brady, M.D., LL.D., D.Sc.,F.R.S. Four plates. 

On the Morphology and Life History of Spirochaeta duttoni . By 

Anton Breinl, M.U.Dr. (Prag.) One plate. 

The Cytology of the Trypanosomes. Part I. By J. E. Salvin-Moore, 
A.R.C.S., F.L.S., F.Z.S., and Anton Breinl, M.U.Dr. (Prag.) Five plates. 

Imp. 8vo. Price 7s. 6d. nett 


ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. I. No. 4. 

Observations on the so-called ‘ Canary Fever.’ By C. E. Walker. 
Contribution k l’dtude de Porocephalus moniliformis. Par A. Broden et 
J. Rodhain. One plate. 

On the Habits, Life-Cycle, and Breeding Places of the Common House- 
Fly (Musca domestica , Linn.). By R. Nkwstead, A.L.S., F.E.S. Six plates. 
Some Notes on the Morphology of Spirochceta duttoni in the Organs of Rats. 

By J. J. van Loghem, M.D., Amsterdam. 

Malaria and History. By W. H. S. Jones, M.A. 

Two New Human Cestodes and a New Linguatulid. By J. W. W. 
Stephens, M.D., Cantab., D.P.H. One plate. 

Imp. 8vo. Price 7s. 6d. nett. 

ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. II. No. 1. 

Atoxyl and Trypanosomiasis. By Sir Rubert Boyce, F.R.S., and Anton 
Breinl, M.U.D. Prag. 

My Experience of Trypanosomiasis in Europeans and its Treatment 
by Atoxyl and other drugs. By Sir Patrick Manson, K.C.M.G., F.R.S. 

Imp. 8vo. Price 7s. 6d. nett. 



ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. II. No. 2. 


Reports of the 4 Sleeping Sickness ’ Expedition to the Zambesi for 
the years 1907-1908. By Allan Kinghorn, M.B., and R. Eustace 
Montgomery, M.R.C.V.S. One map. 

A Report on Trypanosomiasis of Domestic Stock in North-Western 
Rhodesia. By R. Eustace Montgomery, M.R.C.V.S., and Allan 
Kinghorn, M.B. One map and ten charts. 

Report on the Work of the Greek Antimalaria League during the 
year 1907. By M. Hadjimichalis, President, and Jean P. Cardamatis, 
General Secretary. Imp. 8vo. Price 7s. 6d. nett 

ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. II. No. 3 

A Peculiar Intralobular Cirrhosis of the Liver produced by the Protozoal 
Parasite of Kala-azar. By Leonard Rogers, M.D., F.R.C.P., B.S., 
F.R.C.S., I.M.S. One coloured plate. 

What is 4 Schistosomum mansoni ’ Sambon 1907? By Dr. A. Looss. 

The Prevention of Dengue Fever. By E. H. Ross, M.R.C.S. Eng., 
L.R.C.P. Lond. 

The Life History of Trypanosoma lewisi . By J. E. Salvin-Moore, A.R.C.S., 
F.L.S., F.Z.S., Anton Breinl, M.U.Dr. Prag., and Edward Hindle, 
A.R.C.S. Lond. Four plates. 

Notes on the Effects of Therapeutic Agents on Trypanosomes in respect 
to (a) Acquired Resistance of the Parasites to the Drug, and 
( b ) Changes in Virulence of the Strains after Escape from the 
Drug. By Benjamin Moore, M.A., D.Sc. (R.U.I.), Maximilian Nierenstein, 
Ph.D. Berne, and John Lancelot Todd, B.A., M.D. McGill, M.R.C.S. 

Observations on the Acidity and Alkalinity of the Blood in Trypanosome 
Infections. By M. Nierenstein, Ph.D. 

Contributions to the Morphology and Life History of Piroplasma Canis . 
By Anton Breinl, M.U.Dr. Prag., and Edward Hindle, A.R.C.S. Lond. 
Four plates. 

Comparative Chemotherapeutical Study of Atoxyl and Trypanocides. 
Part I. By M. Nierenstein, Ph.D. 

On Three New Species of Culex coPected during the Antimalarial 
Campaign in Mauritius in 1908. M. d’Emmerez de Charmoy. 

Imp. 8vo. Price 7s. 6d. nett. 


vi 



ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. II. No. 4 

Concerning the Treatment of Experimental Trypanosomiasis. Part II. 
By Benjamin Moore, M.A., D.Sc., M. Nierenstein, Ph.D., and John L. 
Todd, M.D. 

An Unusual Case of Goundou. By Dr. R. YV. Orpen. One plate 

Sub-Drainage as Applied to the Anti-Malarial Campaign on the Isthmus 
of Panama. By Henry Simms. 

A New Culicid Genus. By F. V. Theobald, M.A. 

The Inflicted Talipes of the Chinese. By Frank Jeans, M.A., M.B., 
B.C. Cantab., F.R.C.S. Eng. Two plates. 

Contribution & l’6tude de Poroceplialus moniliformis . Par A. Broden et 
J. Rodhain. 

A New Human Nematode, Strongylus gibsoni , n. sp. By J. W. W. Stephens, 
M.D. Cantab. Two plates. 

On the Supposed Occurrence of Filaria immitis in Man. By J. W. W. 
Stephens, M.D. Cantab. 

A New Porocephalus (Poroceplialus cercopitheci y n. sp.). By Anton Breinl, 
M.U.Dr. Prag., and Edward Hindle, A.R.C.S. Lond. 

Comparative Chemo-therapeutical Study of Atoxyl and Trypanocides. 
Part II. By M. Nierenstein, Ph.D. 

Chemical Notes on Atoxyl. By M. Nierenstein, Ph.D. 

Note sur le role des Tabanides dans la Propagation des Trypanosomiases. 
Par Le Dr. Edmond Sergent. Imp. 8vo. Price 7s. 6d. nett. 


ANNALS OF TROPICAL MEDICINE AND 
PARASITOLOGY. Vol. II. No. 5. 

On the Nomenclature of the Mammalian Trypanosomes observed in 
North Western Rhodesia. By R. Eustace Montgomery, M.R.C.V.S., 
and Allan Kinghorn, M.B. Toronto. 

Experiments on the Combined Atoxyl-Mercury Treatment in Monkeys 
Infected with Trypanosoma gambicnsc . By Anton Breinl, M.U.Dr. Prag. 

Drugs from the Congo. By Prosper H. Marsden, F.C.S. 

A Gregarine Parasitic in the Dog-Flea, Ctenocephalus serraticeps. By 
E. H. Ross, M.R.C.S. Eng., L.R.C.P. Lond. 

The Action of Aryl-Stibinic Acids in Experimental Trypanosomiasis. 
By Anton Breinl, M.U.Dr. Prag., and M. Nierenstein, Ph.D. Berne. 

Short Note on the Mechanism of Haemolysis in Piroplasmosis canis . 
By Anton Breinl, M.U.Dr. Prag., and H. E. Annett, M.D., D.P.H. 

Gland Puncture in the Diagnosis of Animal Trypanosomiasis. By 
R. Eustace Montgomery, M.R.C.V.S,, and Allan Kinghorn, M.B. Toronto. 

Observations on the Hooklets of Cysticercus cellulosae in Man. By J. W. W. 
Stephens, M.D. Cantab, D.P.H. Imp. 8vo. Price 7s. 6d. nett. 



TEXT BOOK 


The Practical Study of Malaria and ether Blood 
Parasltee • By J. W. W. Stephens, M D„ Cantab., D.P.H., and S. R. 
Christophers, M.B., Viet., I.M.S. Third edition, thoroughly revised, with six 
coloured plates and one hundred and twenty-eight illustrations in the text. 8vo. 
Price 128. 6d. nett. 


MISCELLANEOUS 

Notes on Sanitary Conditions obtaining in Para (1901). By the 
Members of (1901) The Yellow Fever Expedition. Price is. 

The Habits of the Marine Mosquito. Bv Surgeon Ross, R.N. Price is. 


Published for 

The Committee of the Liverpool School of Tropical 

Medicine 

by 

ARCHIBALD CONSTABLE & CO. 

16 James Street, Haymarket, London, S.W. 

1909 


viii 



LIVERPOOL SCHOOL OF TROPICAL 
MEDICINE 


NOTICE 

Especial attention is called to the fact that the Committee of the 
Liverpool School of Tropical Medicine has decided that the following 
changes shall he made in the courses of instruction. 

i. The Autumn and Lent Courses, which now last only ten weeks,, 
shall be extended to thirteen weeks, followed, as at present, by the 
examination for the Diploma in Tropical Medicine given by the 
University. 


2. In order to allow of this change being made, the present 
Summer Term shall be replaced by a short Course of Practical 
Instruction in Tropical Pathology and Medical Entomology, lasting 
for four weeks in June, and followed by a class examination with 
Certificate of Satisfactory Attendance -the acquisition of this 
Certificate to excuse the first four weeks’ attendance for the full 
Autumn and Lent Courses. 

In accordance with this decision during the next year, 1909, the- 
courses of instruction will be given on the following dates: — 

Full Course begins 6 January. 

Diploma Examination, 5 April. 

Short Course begins 1 June. 

Certificate Examination, 29 June. 

Full Course begins 15 September. 

Diploma Examination, 13 December.